igg3  (Millipore)


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    Name:
    Anti Mouse IgG3 heavy chain specific antibody
    Description:

    Catalog Number:
    m9924
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    Structured Review

    Millipore igg3
    PHB bead production and their immunogenicity. ( a ) Schematic representation of the PHB beads production, composition and immunization schedule. Depicted protein structures are derived from the protein data bank as follows: NadA variant 5 (4CJD), factor H binding protein (mutant G1) (2Y7S). PHB, polyhydroxybutyrate; ( b ) Assessment of antibodies binding to NadA; c ) Assessment of NadA-specific <t>IgG</t> subclass titers using pooled sera from the 6 animals; ( d ) Assessment of antibodies binding to GNA2091-fHbp-G1 specific; ( e ) Assessment of GNA2091-fHbp-G1 specific IgG subclass titers using pooled sera from the 6 animals; ( f ) Serum bactericidal activity (SBA) evaluationis. Statistical significance with ( p

    https://www.bioz.com/result/igg3/product/Millipore
    Average 96 stars, based on 24 article reviews
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    igg3 - by Bioz Stars, 2020-07
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    Images

    1) Product Images from "Bioengineered polyester beads co-displaying protein and carbohydrate-based antigens induce protective immunity against bacterial infection"

    Article Title: Bioengineered polyester beads co-displaying protein and carbohydrate-based antigens induce protective immunity against bacterial infection

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-20205-7

    PHB bead production and their immunogenicity. ( a ) Schematic representation of the PHB beads production, composition and immunization schedule. Depicted protein structures are derived from the protein data bank as follows: NadA variant 5 (4CJD), factor H binding protein (mutant G1) (2Y7S). PHB, polyhydroxybutyrate; ( b ) Assessment of antibodies binding to NadA; c ) Assessment of NadA-specific IgG subclass titers using pooled sera from the 6 animals; ( d ) Assessment of antibodies binding to GNA2091-fHbp-G1 specific; ( e ) Assessment of GNA2091-fHbp-G1 specific IgG subclass titers using pooled sera from the 6 animals; ( f ) Serum bactericidal activity (SBA) evaluationis. Statistical significance with ( p
    Figure Legend Snippet: PHB bead production and their immunogenicity. ( a ) Schematic representation of the PHB beads production, composition and immunization schedule. Depicted protein structures are derived from the protein data bank as follows: NadA variant 5 (4CJD), factor H binding protein (mutant G1) (2Y7S). PHB, polyhydroxybutyrate; ( b ) Assessment of antibodies binding to NadA; c ) Assessment of NadA-specific IgG subclass titers using pooled sera from the 6 animals; ( d ) Assessment of antibodies binding to GNA2091-fHbp-G1 specific; ( e ) Assessment of GNA2091-fHbp-G1 specific IgG subclass titers using pooled sera from the 6 animals; ( f ) Serum bactericidal activity (SBA) evaluationis. Statistical significance with ( p

    Techniques Used: Derivative Assay, Variant Assay, Binding Assay, Mutagenesis, Activity Assay

    Bactericidal activity of various sera. ( A ) N. meningitidis serogroup C (Strain C11); ( B ) N. meningitidis serogroup B (Strain CU385/83); ( C ) N. meningitidis serogroup A (Strain 499/03). Bactericidal titers are expressed as mean ± SEM (8 mice each group) of the reciprocal value of the greatest sera dilution leading to ≥50% of killing. Statistical significant were determinate by an ANOVA by Kruskal-Wallis test with Dunn’s multiple comparison post-test in Fig. 6A and labelled with letters. Groups MenC-PhaC presented superior IgG levels than group MenC-NadA-PhaC, MenC-NadA-His6 and MenC-TD with a, b ( p
    Figure Legend Snippet: Bactericidal activity of various sera. ( A ) N. meningitidis serogroup C (Strain C11); ( B ) N. meningitidis serogroup B (Strain CU385/83); ( C ) N. meningitidis serogroup A (Strain 499/03). Bactericidal titers are expressed as mean ± SEM (8 mice each group) of the reciprocal value of the greatest sera dilution leading to ≥50% of killing. Statistical significant were determinate by an ANOVA by Kruskal-Wallis test with Dunn’s multiple comparison post-test in Fig. 6A and labelled with letters. Groups MenC-PhaC presented superior IgG levels than group MenC-NadA-PhaC, MenC-NadA-His6 and MenC-TD with a, b ( p

    Techniques Used: Activity Assay, Mouse Assay

    2) Product Images from "Combining Monophosphoryl Lipid A (MPL), CpG Oligodeoxynucleotide (ODN), and QS-21 Adjuvants Induces Strong and Persistent Functional Antibodies and T Cell Responses against Cell-Traversal Protein for Ookinetes and Sporozoites (CelTOS) of Plasmodium falciparum in BALB/c Mice"

    Article Title: Combining Monophosphoryl Lipid A (MPL), CpG Oligodeoxynucleotide (ODN), and QS-21 Adjuvants Induces Strong and Persistent Functional Antibodies and T Cell Responses against Cell-Traversal Protein for Ookinetes and Sporozoites (CelTOS) of Plasmodium falciparum in BALB/c Mice

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00911-18

    Evaluation of endpoint titers of anti-PfCelTOS total IgG, IgG2a, and IgG2b antibodies. For titration, 1:200 to 1:409,600 dilutions of mouse sera from different vaccine groups (groups 1 to 5) were analyzed using an ELISA. The bars show the last dilution
    Figure Legend Snippet: Evaluation of endpoint titers of anti-PfCelTOS total IgG, IgG2a, and IgG2b antibodies. For titration, 1:200 to 1:409,600 dilutions of mouse sera from different vaccine groups (groups 1 to 5) were analyzed using an ELISA. The bars show the last dilution

    Techniques Used: Titration, Enzyme-linked Immunosorbent Assay

    Anti-PfCelTOS IgG antibody responses and persistence of IgG responses.
    Figure Legend Snippet: Anti-PfCelTOS IgG antibody responses and persistence of IgG responses.

    Techniques Used:

    Avidity analyses of anti-PfCelTOS IgG, IgG2a, and IgG2b antibodies by an ELISA. The avidity index (AI) was calculated as the portion of the OD value of urea-treated serum samples compared to that of untreated samples multiplied by 100. AI values of
    Figure Legend Snippet: Avidity analyses of anti-PfCelTOS IgG, IgG2a, and IgG2b antibodies by an ELISA. The avidity index (AI) was calculated as the portion of the OD value of urea-treated serum samples compared to that of untreated samples multiplied by 100. AI values of

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Assessment of anti-PfCelTOS IgG subclass profiles and longevity. Preimmune sera ( n  = 30) were used as negative controls to determine the ELISA cutoffs, which were calculated as the mean OD 490 plus 3 SD. The cutoffs for total IgG (TIgG),
    Figure Legend Snippet: Assessment of anti-PfCelTOS IgG subclass profiles and longevity. Preimmune sera ( n  = 30) were used as negative controls to determine the ELISA cutoffs, which were calculated as the mean OD 490 plus 3 SD. The cutoffs for total IgG (TIgG),

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Anti-PfCelTOS IgG antibody levels in vaccine mouse sera at different time points by an ELISA. Groups of 6- to 8-week-old female BALB/c mice were immunized subcutaneously with recombinant PfCelTOS alone or formulated with different adjuvants (MPL, CpG,
    Figure Legend Snippet: Anti-PfCelTOS IgG antibody levels in vaccine mouse sera at different time points by an ELISA. Groups of 6- to 8-week-old female BALB/c mice were immunized subcutaneously with recombinant PfCelTOS alone or formulated with different adjuvants (MPL, CpG,

    Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay, Recombinant

    Evaluation of Th1/Th2 ratios. Th1 (anti-PfCelTOS IgG2a and IgG2b antibodies and cytokines IFN-γ and TNF-α) and Th2 (anti-PfCelTOS IgG1 and the cytokine IL-10) responses were analyzed in different vaccine groups on days 38 and 180 after
    Figure Legend Snippet: Evaluation of Th1/Th2 ratios. Th1 (anti-PfCelTOS IgG2a and IgG2b antibodies and cytokines IFN-γ and TNF-α) and Th2 (anti-PfCelTOS IgG1 and the cytokine IL-10) responses were analyzed in different vaccine groups on days 38 and 180 after

    Techniques Used:

    3) Product Images from "The invA gene of Brucella melitensis is involved in intracellular invasion and is required to establish infection in a mouse model"

    Article Title: The invA gene of Brucella melitensis is involved in intracellular invasion and is required to establish infection in a mouse model

    Journal: Virulence

    doi: 10.4161/viru.28589

    Figure 6. Immune responses of infected mouse groups. ( A–E ) correspond to the humoral response. Results are expressed as the mean O.D. 490 ± standard deviation at different times pi. ( A ) IgG1 antibody results, ( B ) IgG2a, ( C ) IgG2b,
    Figure Legend Snippet: Figure 6. Immune responses of infected mouse groups. ( A–E ) correspond to the humoral response. Results are expressed as the mean O.D. 490 ± standard deviation at different times pi. ( A ) IgG1 antibody results, ( B ) IgG2a, ( C ) IgG2b,

    Techniques Used: Infection, Standard Deviation

    4) Product Images from "IgG4 autoantibodies induce dermal–epidermal separation"

    Article Title: IgG4 autoantibodies induce dermal–epidermal separation

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/j.1582-4934.2007.00081.x

    Purification of IgG1 and IgG4 autoantibodies specific to the epidermal basement membrane. IF microscopy study of serum from patient 1 on normal human skin demonstrates deposition of mainly IgG1 (A) and IgG4 (D) and traces of IgG2 (B) and IgG3 (C) at the dermal–epidermal junction. After purification against an affinity matrix containing mon-oclonal antibodies specific for IgG1, IgG1 autoantibodies to the skin basement membrane can be visualized in the purified fraction (E) , while IgG2 (F) , IgG3 (G) and IgG4 (H) autoantibodies are not detectable. Conversely, the antibody fraction purified against an affinity matrix specific for IgG4 demonstrated binding of autoantibodies to the basement membrane of normal human skin exclusively belonging to the IgG4 subclass (L) , whereas no deposits of IgG1 (I) , IgG2 (J) or IgG3 (K) were observed (all magnifications, ×200).
    Figure Legend Snippet: Purification of IgG1 and IgG4 autoantibodies specific to the epidermal basement membrane. IF microscopy study of serum from patient 1 on normal human skin demonstrates deposition of mainly IgG1 (A) and IgG4 (D) and traces of IgG2 (B) and IgG3 (C) at the dermal–epidermal junction. After purification against an affinity matrix containing mon-oclonal antibodies specific for IgG1, IgG1 autoantibodies to the skin basement membrane can be visualized in the purified fraction (E) , while IgG2 (F) , IgG3 (G) and IgG4 (H) autoantibodies are not detectable. Conversely, the antibody fraction purified against an affinity matrix specific for IgG4 demonstrated binding of autoantibodies to the basement membrane of normal human skin exclusively belonging to the IgG4 subclass (L) , whereas no deposits of IgG1 (I) , IgG2 (J) or IgG3 (K) were observed (all magnifications, ×200).

    Techniques Used: Purification, Microscopy, Binding Assay

    A blocking monoclonal antibody against CD16 significantly inhibits the autoantibody-induced dermal–epidermal separation. Cryo-sections of human skin were incubated with IgG, IgG1 and IgG4 purified from BP patients 1 and 2 (n = 4 sections/antibody preparation). Subsequently, granulocytes were treated for 10 min at room temperature with 3G8 or a control antibody prior to incubation with skin sections. Dermal–epidermal separation is represented as means ± SEM of the percent of the total length of the dermal–epidermal junction (DEJ) for each section.
    Figure Legend Snippet: A blocking monoclonal antibody against CD16 significantly inhibits the autoantibody-induced dermal–epidermal separation. Cryo-sections of human skin were incubated with IgG, IgG1 and IgG4 purified from BP patients 1 and 2 (n = 4 sections/antibody preparation). Subsequently, granulocytes were treated for 10 min at room temperature with 3G8 or a control antibody prior to incubation with skin sections. Dermal–epidermal separation is represented as means ± SEM of the percent of the total length of the dermal–epidermal junction (DEJ) for each section.

    Techniques Used: Blocking Assay, Incubation, Purification

    IgG4 autoantibodies, in contrast to IgG1, do not fix complement to the dermal–epidermal junction. In a representative experiment, cryosections of normal human skin were incubated with serum and purified antibody preparations from patient 2 and, subsequently, treated with fresh human serum as a source of complement. Both serum (A) and purified IgG1 autoantibodies (B) fixed complement C3 at the dermal–epidermal junction in a linear fashion. In contrast, incubation of cryosections with IgG4 specific for the dermal–epidermal junction (C) or serum from a healthy control (D) does not result in C3 deposition (all magnifications, ×200).
    Figure Legend Snippet: IgG4 autoantibodies, in contrast to IgG1, do not fix complement to the dermal–epidermal junction. In a representative experiment, cryosections of normal human skin were incubated with serum and purified antibody preparations from patient 2 and, subsequently, treated with fresh human serum as a source of complement. Both serum (A) and purified IgG1 autoantibodies (B) fixed complement C3 at the dermal–epidermal junction in a linear fashion. In contrast, incubation of cryosections with IgG4 specific for the dermal–epidermal junction (C) or serum from a healthy control (D) does not result in C3 deposition (all magnifications, ×200).

    Techniques Used: Incubation, Purification

    IgG4 autoantibodies recruit and activate leucocytes. Sections of human skin were incubated with IgG1 and IgG4 autoantibodies from a BP patient, as well as with IgG from a healthy control. Subsequent addition of leucocytes from healthy donors leads to leucocyte attachment at the dermal–epidermal junction in sections treated with patient's IgG1 (A) and IgG4 autoantibodies (B) , but not in sections incubated with control IgG (C) . Activation of leucocytes, as revealed by the reduction of nitro blue tetrazolium (NBT) to formazan (dark precipitates), is induced by purified IgG1 (D) and IgG4 autoantibodies (E) , but not by control IgG (F) (all magnifications, ×400). (G) Cryosections of human skin were treated with IgG1 (n = 3) and IgG4 (n = 5) antibodies (four sections/antibody preparation). Subsequently, leucocytes from healthy donors were incubated for 90 min with the cryosections. Deposition of formazan is represented as means ± SEM of the percent of the total length of the dermal–epidermal junction (DEJ) for each section.
    Figure Legend Snippet: IgG4 autoantibodies recruit and activate leucocytes. Sections of human skin were incubated with IgG1 and IgG4 autoantibodies from a BP patient, as well as with IgG from a healthy control. Subsequent addition of leucocytes from healthy donors leads to leucocyte attachment at the dermal–epidermal junction in sections treated with patient's IgG1 (A) and IgG4 autoantibodies (B) , but not in sections incubated with control IgG (C) . Activation of leucocytes, as revealed by the reduction of nitro blue tetrazolium (NBT) to formazan (dark precipitates), is induced by purified IgG1 (D) and IgG4 autoantibodies (E) , but not by control IgG (F) (all magnifications, ×400). (G) Cryosections of human skin were treated with IgG1 (n = 3) and IgG4 (n = 5) antibodies (four sections/antibody preparation). Subsequently, leucocytes from healthy donors were incubated for 90 min with the cryosections. Deposition of formazan is represented as means ± SEM of the percent of the total length of the dermal–epidermal junction (DEJ) for each section.

    Techniques Used: Incubation, Activation Assay, Purification

    IgG4 autoantibodies show a lower pathogenic capacity compared with IgG1. In the upper panel, cryosections of human skin were incubated with IgG1 or IgG4 autoantibodies purified from patient 2, both adjusted at an end-point titer of 1:80 by IF microscopy. Subsequent addition of leucocytes leads to blister formation in cryosections treated with IgG1 autoantibodies (A) . In contrast, IgG4 autoantibodies (B) fail to recruit leucocytes to the dermal–epidermal junction and to induce dermal–epidermal separation (magnification, ×400). The lower panel (C) shows the extent of dermal–epidermal separation after incubating the cryosections with IgG1 and IgG4 preparations from two BP patients (n = 4 sections/antibody preparation). Dermal–epidermal separation is represented as means ± SEM of the percent of the total length of the dermal–epidermal junction (DEJ) for each section. The reactivity of the antibody preparations as determined by their end-point titres by immunofluorescence microscopy is represented on the X-axis.
    Figure Legend Snippet: IgG4 autoantibodies show a lower pathogenic capacity compared with IgG1. In the upper panel, cryosections of human skin were incubated with IgG1 or IgG4 autoantibodies purified from patient 2, both adjusted at an end-point titer of 1:80 by IF microscopy. Subsequent addition of leucocytes leads to blister formation in cryosections treated with IgG1 autoantibodies (A) . In contrast, IgG4 autoantibodies (B) fail to recruit leucocytes to the dermal–epidermal junction and to induce dermal–epidermal separation (magnification, ×400). The lower panel (C) shows the extent of dermal–epidermal separation after incubating the cryosections with IgG1 and IgG4 preparations from two BP patients (n = 4 sections/antibody preparation). Dermal–epidermal separation is represented as means ± SEM of the percent of the total length of the dermal–epidermal junction (DEJ) for each section. The reactivity of the antibody preparations as determined by their end-point titres by immunofluorescence microscopy is represented on the X-axis.

    Techniques Used: Incubation, Purification, Microscopy, Immunofluorescence

    IgG4 autoantibodies induce dermal–epidermal separation in sections of human skin. Results of a representative experiment show that dermal–epidermal separation in sections of normal human skin is induced by serum (A) , IgG1 (B) and IgG4 autoantibodies ( C ) from patient 1. Serum antibodies from a healthy control (D) do not induce sub-epidermal splits (all magnifications, ×200).
    Figure Legend Snippet: IgG4 autoantibodies induce dermal–epidermal separation in sections of human skin. Results of a representative experiment show that dermal–epidermal separation in sections of normal human skin is induced by serum (A) , IgG1 (B) and IgG4 autoantibodies ( C ) from patient 1. Serum antibodies from a healthy control (D) do not induce sub-epidermal splits (all magnifications, ×200).

    Techniques Used:

    5) Product Images from "Immunologic Responses to Vibrio cholerae in Patients Co-Infected with Intestinal Parasites in Bangladesh"

    Article Title: Immunologic Responses to Vibrio cholerae in Patients Co-Infected with Intestinal Parasites in Bangladesh

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0000403

    CTB specific IgA and IgG responses are shown in cholera patients across classes of helminth infection (A and B, respectively). The hatched bars indicate the mean log2 transformed CTB titer on day 2, and the dark bars indicate the titer on day 21 and error bars indicate the standard error of the mean. An asterisk denotes a statistically significant difference between non-infected and infected individuals. All classes of helminth infected patients had significantly lower CTB IgA immune responses on day 21 compared to non-helminth infected controls.
    Figure Legend Snippet: CTB specific IgA and IgG responses are shown in cholera patients across classes of helminth infection (A and B, respectively). The hatched bars indicate the mean log2 transformed CTB titer on day 2, and the dark bars indicate the titer on day 21 and error bars indicate the standard error of the mean. An asterisk denotes a statistically significant difference between non-infected and infected individuals. All classes of helminth infected patients had significantly lower CTB IgA immune responses on day 21 compared to non-helminth infected controls.

    Techniques Used: CtB Assay, Infection, Transformation Assay

    6) Product Images from "Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation"

    Article Title: Development of Immunocapture-LC/MS Assay for Simultaneous ADA Isotyping and Semiquantitation

    Journal: Journal of Immunology Research

    doi: 10.1155/2016/7682472

    Calibration curves of IgG1 in human plasma eluent after immunocapture when using either drug (a) or mouse mAb as ADA capture reagent (b).
    Figure Legend Snippet: Calibration curves of IgG1 in human plasma eluent after immunocapture when using either drug (a) or mouse mAb as ADA capture reagent (b).

    Techniques Used:

    LC/MS chromatograms of unique peptides of IgG1 (top), IgM (middle), and IgE (bottom) from blank human plasma (left) and LLOQ samples (right) after immunocapture when using mouse mAb for ADA capture.
    Figure Legend Snippet: LC/MS chromatograms of unique peptides of IgG1 (top), IgM (middle), and IgE (bottom) from blank human plasma (left) and LLOQ samples (right) after immunocapture when using mouse mAb for ADA capture.

    Techniques Used: Liquid Chromatography with Mass Spectroscopy

    LC/MS chromatograms of unique peptides of IgG1 (top), IgM (middle), and IgE (bottom) from blank human plasma (left) and LLOQ samples (right) after immunocapture when using drug as ADA capture reagent.
    Figure Legend Snippet: LC/MS chromatograms of unique peptides of IgG1 (top), IgM (middle), and IgE (bottom) from blank human plasma (left) and LLOQ samples (right) after immunocapture when using drug as ADA capture reagent.

    Techniques Used: Liquid Chromatography with Mass Spectroscopy

    7) Product Images from "An evolutionary approach to identify potentially protective B cell epitopes involved in naturally acquired immunity to malaria and the role of EBA-175 in protection amongst denizens of Bolifamba, Cameroon"

    Article Title: An evolutionary approach to identify potentially protective B cell epitopes involved in naturally acquired immunity to malaria and the role of EBA-175 in protection amongst denizens of Bolifamba, Cameroon

    Journal: Malaria Journal

    doi: 10.1186/s12936-016-1337-z

    Mean optical densities (ODs) for IgG3 antibody subclass to; a EBA-175, b PF4-123, c PF4-143. The bars represent the arithmetic mean OD values of European sera (ES), healthy children (HC), sick children (SC), sick adults (SA), and healthy adults (HA)
    Figure Legend Snippet: Mean optical densities (ODs) for IgG3 antibody subclass to; a EBA-175, b PF4-123, c PF4-143. The bars represent the arithmetic mean OD values of European sera (ES), healthy children (HC), sick children (SC), sick adults (SA), and healthy adults (HA)

    Techniques Used:

    Correlation and regression equation of parasite load to total IgG antibodies to; a EBA-175, b PF4-123, c PF4-143. r Spearman’s rank correlation coefficient, p two-tailed level of significance
    Figure Legend Snippet: Correlation and regression equation of parasite load to total IgG antibodies to; a EBA-175, b PF4-123, c PF4-143. r Spearman’s rank correlation coefficient, p two-tailed level of significance

    Techniques Used: Two Tailed Test

    Correlation and regression equation of duration of stay in Bolifamba to parasite load ( a ) and total IgG antibodies to; ( b ) EBA-175, ( c ) PF4-123, ( d ) PF4-143. r Spearman’s rank correlation coefficient, p two-tailed level of significance
    Figure Legend Snippet: Correlation and regression equation of duration of stay in Bolifamba to parasite load ( a ) and total IgG antibodies to; ( b ) EBA-175, ( c ) PF4-123, ( d ) PF4-143. r Spearman’s rank correlation coefficient, p two-tailed level of significance

    Techniques Used: Two Tailed Test

    Mean optical densities (ODs) for IgG1 antibody subclass to; a EBA-175, b PF4-123, c PF4-143. The bars represent the arithmetic mean OD values of European sera (ES), healthy children (HC), sick children (SC), sick adults (SA), and healthy adults (HA)
    Figure Legend Snippet: Mean optical densities (ODs) for IgG1 antibody subclass to; a EBA-175, b PF4-123, c PF4-143. The bars represent the arithmetic mean OD values of European sera (ES), healthy children (HC), sick children (SC), sick adults (SA), and healthy adults (HA)

    Techniques Used:

    Mean optical densities (ODs) for total IgG; a EBA-175, b PF4-123, c PF4-143. The bars represent the arithmetic mean OD values of European sera (ES), healthy children (HC), sick children (SC), sick adults (SA), and healthy adults (HA). More details are under materials and methods
    Figure Legend Snippet: Mean optical densities (ODs) for total IgG; a EBA-175, b PF4-123, c PF4-143. The bars represent the arithmetic mean OD values of European sera (ES), healthy children (HC), sick children (SC), sick adults (SA), and healthy adults (HA). More details are under materials and methods

    Techniques Used:

    8) Product Images from "Membranous Nephropathy Associated With Immunological Disorder-Related Liver Disease"

    Article Title: Membranous Nephropathy Associated With Immunological Disorder-Related Liver Disease

    Journal: Medicine

    doi: 10.1097/MD.0000000000001243

    Immunofluorescence staining for IgG subclasses in patient 8 with PSC and AIH (original magnification ×400). Granular staining for (A) IgG1 and (D) IgG4 in the subepithelial portion along the glomerular capillary wall with predominance of IgG4. Immunofluorescence staining for (B) IgG2 and (C) IgG3 was negative. AIH = autoimmune hepatitis, IgG = immunoglobulin G, PSC = primary sclerosing cholangitis.
    Figure Legend Snippet: Immunofluorescence staining for IgG subclasses in patient 8 with PSC and AIH (original magnification ×400). Granular staining for (A) IgG1 and (D) IgG4 in the subepithelial portion along the glomerular capillary wall with predominance of IgG4. Immunofluorescence staining for (B) IgG2 and (C) IgG3 was negative. AIH = autoimmune hepatitis, IgG = immunoglobulin G, PSC = primary sclerosing cholangitis.

    Techniques Used: Immunofluorescence, Staining

    9) Product Images from "Lack of Humoral Immune Protection against Treponema denticola Virulence in a Murine Model"

    Article Title: Lack of Humoral Immune Protection against Treponema denticola Virulence in a Murine Model

    Journal: Infection and Immunity

    doi:

    (Left) Ability of serum antibody incubated with T. denticola in vitro to alter lesion development following reinfection. The bars indicate means for five animals per group challenged with 10 10 T. denticola organisms after incubation with primary-infection sera and rabbit complement (Primary/C′) for 1 or 3 h, with control normal mouse serum and complement for 3 h, or with complement alone for 3 h. Td indicates control T. denticola maintained in NOS medium for 3 h (similar to the experimental incubations). The error bars indicate one standard deviation. (Right) Serum IgG antibody levels in mice challenged with T. denticola following in vitro incubations as described above. The bars indicate the mean serum IgG levels of 5 to 10 mice per group, and the error bars indicate one standard deviation. The paired symbols indicate IgG antibody levels significantly different at P
    Figure Legend Snippet: (Left) Ability of serum antibody incubated with T. denticola in vitro to alter lesion development following reinfection. The bars indicate means for five animals per group challenged with 10 10 T. denticola organisms after incubation with primary-infection sera and rabbit complement (Primary/C′) for 1 or 3 h, with control normal mouse serum and complement for 3 h, or with complement alone for 3 h. Td indicates control T. denticola maintained in NOS medium for 3 h (similar to the experimental incubations). The error bars indicate one standard deviation. (Right) Serum IgG antibody levels in mice challenged with T. denticola following in vitro incubations as described above. The bars indicate the mean serum IgG levels of 5 to 10 mice per group, and the error bars indicate one standard deviation. The paired symbols indicate IgG antibody levels significantly different at P

    Techniques Used: Incubation, In Vitro, Infection, Standard Deviation, Mouse Assay

    (Left) Serum IgG subclass antibody levels expressed as ELISA units, following primary infection or reinfection with T. denticola . The bars indicate the means from 10 to 15 animals derived from three independent experiments, and the error bars indicate one standard deviation. ∗, significantly less than all other subclasses at P
    Figure Legend Snippet: (Left) Serum IgG subclass antibody levels expressed as ELISA units, following primary infection or reinfection with T. denticola . The bars indicate the means from 10 to 15 animals derived from three independent experiments, and the error bars indicate one standard deviation. ∗, significantly less than all other subclasses at P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Derivative Assay, Standard Deviation

    10) Product Images from "Long-term renal survival of γ3-heavy chain deposition disease: a case report"

    Article Title: Long-term renal survival of γ3-heavy chain deposition disease: a case report

    Journal: BMC Nephrology

    doi: 10.1186/s12882-017-0645-z

    Immunofluorescence staining shows moderate positivity for IgG ( a ) and C3 ( b ) within the mesangial nodules and in a linear deposition along the capillary walls. However, κ ( c ) and λ ( d ) are undetectable in the same glomerule. Original magnification 400×. e-h Immunofluorescence findings for IgG subclasses. Only IgG3 ( g ) is intensely positive in the mesangium area and along the glomerular and tubular basement membranes. Staining for IgG1 ( e ), IgG2 ( f ), and IgG4 ( h ) was absolutely negative in the same specimen. Original magnification 100×
    Figure Legend Snippet: Immunofluorescence staining shows moderate positivity for IgG ( a ) and C3 ( b ) within the mesangial nodules and in a linear deposition along the capillary walls. However, κ ( c ) and λ ( d ) are undetectable in the same glomerule. Original magnification 400×. e-h Immunofluorescence findings for IgG subclasses. Only IgG3 ( g ) is intensely positive in the mesangium area and along the glomerular and tubular basement membranes. Staining for IgG1 ( e ), IgG2 ( f ), and IgG4 ( h ) was absolutely negative in the same specimen. Original magnification 100×

    Techniques Used: Immunofluorescence, Staining

    11) Product Images from "Binding to histo-blood group antigen-expressing bacteria protects human norovirus from acute heat stress"

    Article Title: Binding to histo-blood group antigen-expressing bacteria protects human norovirus from acute heat stress

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2015.00659

    Examples of flow cytometry and fluorescence microscopy results. (A) Flow cytometry plot of Escherichia coli LMG8223 stained by an irrelevant IgG3 (negative control); (B) Flow cytometry plot of E. coli LMG8223 stained by anti-A antibody #21; (C) Fluorescence microscopy picture of E. coli LMG8223 stained with an irrelevant IgG3 (negative control); (D) Fluorescence microscopy picture of E. coli LMG8223 stained by anti-A antibody #21.
    Figure Legend Snippet: Examples of flow cytometry and fluorescence microscopy results. (A) Flow cytometry plot of Escherichia coli LMG8223 stained by an irrelevant IgG3 (negative control); (B) Flow cytometry plot of E. coli LMG8223 stained by anti-A antibody #21; (C) Fluorescence microscopy picture of E. coli LMG8223 stained with an irrelevant IgG3 (negative control); (D) Fluorescence microscopy picture of E. coli LMG8223 stained by anti-A antibody #21.

    Techniques Used: Flow Cytometry, Cytometry, Fluorescence, Microscopy, Staining, Negative Control

    12) Product Images from "Malaria vaccine candidate based on Duffy-binding protein elicits strain transcending functional antibodies in a Phase I trial"

    Article Title: Malaria vaccine candidate based on Duffy-binding protein elicits strain transcending functional antibodies in a Phase I trial

    Journal: NPJ Vaccines

    doi: 10.1038/s41541-018-0083-3

    Humoral response to immunization with PvDBPII formulated with GLA-SE. a Kinetics of PvDBPII-specific Geometric Mean antibody titres for all Study Groups (PP Population). Geometric mean antibody levels (U/mL) to PvDBPII measured by ELISA in sera collected from groups immunized with 10, 25, or 50 μg PvDBPII/GLA-SE and Hepatitis B vaccine. Study participants were immunized on Days 0, 28 and 56, and sera were collected on Days 0, 28, 56, 84 and 180. Antibody levels measured by ELISA were expressed as Geometric Mean Titres (GMT) with 95% confidence interval for the nine subjects. b Serum IgG subclass responses to PvDBPII in three PvDBPII dose groups (PP Population). Geometric means with SD are shown
    Figure Legend Snippet: Humoral response to immunization with PvDBPII formulated with GLA-SE. a Kinetics of PvDBPII-specific Geometric Mean antibody titres for all Study Groups (PP Population). Geometric mean antibody levels (U/mL) to PvDBPII measured by ELISA in sera collected from groups immunized with 10, 25, or 50 μg PvDBPII/GLA-SE and Hepatitis B vaccine. Study participants were immunized on Days 0, 28 and 56, and sera were collected on Days 0, 28, 56, 84 and 180. Antibody levels measured by ELISA were expressed as Geometric Mean Titres (GMT) with 95% confidence interval for the nine subjects. b Serum IgG subclass responses to PvDBPII in three PvDBPII dose groups (PP Population). Geometric means with SD are shown

    Techniques Used: Enzyme-linked Immunosorbent Assay

    13) Product Images from "Identification of the flotillin-1/2 heterocomplex as a target of autoantibodies in bona fide multiple sclerosis"

    Article Title: Identification of the flotillin-1/2 heterocomplex as a target of autoantibodies in bona fide multiple sclerosis

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-017-0900-z

    Histo-immunoprecipitation and antigen identification. Cryosections of rat or pig cerebellum were incubated with the serum (1:100), washed in PBS, and solubilized using detergents. The solution was incubated with protein-G-coated magnetic beads. The immunocomplexes were eluted by SDS and subjected to SDS-PAGE analysis and Western blot. A Western blot after incubation with anti-flotillin-2 ( A1 ) and enzymatic visualization of antibody binding. Staining of SDS polyacrylamide gel with colloidal Coomassie ( A2 ). Lane 1: molecular mass (kDa) marker; lanes 2–8: histo-immunoprecipitates of patient sera from rat cerebellum; lanes 9–15: histo-immunoprecipitates of control samples. The arrow indicates the position of the immunoprecipitated antigen 50 kDa while dotted arrows indicate the position of IgG heavy and light chain at 52 and 27 kDa, respectively. PS patient sample, CS control sample. B Immunofluorescence staining of rat hippocampus ( B1 ) and cerebellum ( B2 ) and primate cerebellum ( B3 ) tissue sections with patient serum ( green , 1–3 ) and anti-flotillin-2 antibody ( red , 1′–3′ ). The merged images show localization of the reactivity in the same region including the more intense staining of the sm internum on the hippocampus ( 1″–3″ ). Scale bar : 50 μm (large images), 100 μm (inserts). PS patient serum, CS control serum, h hilus, sg stratum granulosum, sm stratum moleculare, smi stratum moleculare internum, sme stratum moleculare externum, sp stratum purkinjense
    Figure Legend Snippet: Histo-immunoprecipitation and antigen identification. Cryosections of rat or pig cerebellum were incubated with the serum (1:100), washed in PBS, and solubilized using detergents. The solution was incubated with protein-G-coated magnetic beads. The immunocomplexes were eluted by SDS and subjected to SDS-PAGE analysis and Western blot. A Western blot after incubation with anti-flotillin-2 ( A1 ) and enzymatic visualization of antibody binding. Staining of SDS polyacrylamide gel with colloidal Coomassie ( A2 ). Lane 1: molecular mass (kDa) marker; lanes 2–8: histo-immunoprecipitates of patient sera from rat cerebellum; lanes 9–15: histo-immunoprecipitates of control samples. The arrow indicates the position of the immunoprecipitated antigen 50 kDa while dotted arrows indicate the position of IgG heavy and light chain at 52 and 27 kDa, respectively. PS patient sample, CS control sample. B Immunofluorescence staining of rat hippocampus ( B1 ) and cerebellum ( B2 ) and primate cerebellum ( B3 ) tissue sections with patient serum ( green , 1–3 ) and anti-flotillin-2 antibody ( red , 1′–3′ ). The merged images show localization of the reactivity in the same region including the more intense staining of the sm internum on the hippocampus ( 1″–3″ ). Scale bar : 50 μm (large images), 100 μm (inserts). PS patient serum, CS control serum, h hilus, sg stratum granulosum, sm stratum moleculare, smi stratum moleculare internum, sme stratum moleculare externum, sp stratum purkinjense

    Techniques Used: Immunoprecipitation, Incubation, Magnetic Beads, SDS Page, Western Blot, Binding Assay, Staining, Marker, Immunofluorescence

    Immunofluorescence staining of central nervous tissues. Cryosections of rat hippocampus ( A ) and cerebellum ( B ) as well as primate cerebellum ( C ) were incubated with patient serum ( A – C ) or with control serum ( A′–C′ ) ( A – C1 , A′–C′ 1:100, A – C2 , C3 1:50) in the first step and with Alexa Fluor 488-labeled goat anti-human IgG in the second step ( green ). Nuclei were counterstained by incubation with TO-PRO-3 iodide ( blue ). A fine-granular staining of the stratum moleculare ( sm ) was obtained. On the hippocampus, the sm internum was stained more intensely than the sm externum. Scale bar : 100 μm ( A – C1 , A′–C′ ), 20 μm ( A – C2, C3 ). h hilus, sm stratum moleculare, smi stratum moleculare internum, sme stratum moleculare externum, sg stratum granulosum, sp stratum purkinjense
    Figure Legend Snippet: Immunofluorescence staining of central nervous tissues. Cryosections of rat hippocampus ( A ) and cerebellum ( B ) as well as primate cerebellum ( C ) were incubated with patient serum ( A – C ) or with control serum ( A′–C′ ) ( A – C1 , A′–C′ 1:100, A – C2 , C3 1:50) in the first step and with Alexa Fluor 488-labeled goat anti-human IgG in the second step ( green ). Nuclei were counterstained by incubation with TO-PRO-3 iodide ( blue ). A fine-granular staining of the stratum moleculare ( sm ) was obtained. On the hippocampus, the sm internum was stained more intensely than the sm externum. Scale bar : 100 μm ( A – C1 , A′–C′ ), 20 μm ( A – C2, C3 ). h hilus, sm stratum moleculare, smi stratum moleculare internum, sme stratum moleculare externum, sg stratum granulosum, sp stratum purkinjense

    Techniques Used: Immunofluorescence, Staining, Incubation, Labeling

    14) Product Images from "Characterization of the Antibody Response to the Saliva of Phlebotomus papatasi in People Living in Endemic Areas of Cutaneous Leishmaniasis"

    Article Title: Characterization of the Antibody Response to the Saliva of Phlebotomus papatasi in People Living in Endemic Areas of Cutaneous Leishmaniasis

    Journal: The American Journal of Tropical Medicine and Hygiene

    doi: 10.4269/ajtmh.2011.10-0598

    IgG subclasses and IgE immunoblots against salivary proteins of Phlebotomus papatasi . Salivary gland proteins of P. papatasi were fractioned with a 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). IgG subclasses and IgE antibodies
    Figure Legend Snippet: IgG subclasses and IgE immunoblots against salivary proteins of Phlebotomus papatasi . Salivary gland proteins of P. papatasi were fractioned with a 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). IgG subclasses and IgE antibodies

    Techniques Used: Western Blot, Polyacrylamide Gel Electrophoresis, SDS Page

    Quantification of IgG subclasses and IgE anti-saliva antibodies. Levels of IgG1, IgG2, IgG3, IgG4, and IgE antibodies directed against Phlebotomus papatasi saliva were studied in serum samples of 65 participants with specific IgG antibodies. ( A ) The percentage
    Figure Legend Snippet: Quantification of IgG subclasses and IgE anti-saliva antibodies. Levels of IgG1, IgG2, IgG3, IgG4, and IgE antibodies directed against Phlebotomus papatasi saliva were studied in serum samples of 65 participants with specific IgG antibodies. ( A ) The percentage

    Techniques Used:

    Serum levels of IgG antibodies to Phlebotomus papatasi saliva in people living in endemic areas of Leishmania major transmission. IgG antibodies to salivary gland extracts from P. papatasi were investigated in 200 children who live in endemic areas of
    Figure Legend Snippet: Serum levels of IgG antibodies to Phlebotomus papatasi saliva in people living in endemic areas of Leishmania major transmission. IgG antibodies to salivary gland extracts from P. papatasi were investigated in 200 children who live in endemic areas of

    Techniques Used: Transmission Assay

    15) Product Images from "Biochemical Characterization and Evaluation of a Brugia malayi Small Heat Shock Protein as a Vaccine against Lymphatic Filariasis"

    Article Title: Biochemical Characterization and Evaluation of a Brugia malayi Small Heat Shock Protein as a Vaccine against Lymphatic Filariasis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0034077

    Isotype of anti-BmHsp12.6 IgG antibodies in the sera of mice. Mice were immunized with A ) BmHsp12.6, B ) BmHsp12.6αc or C ) N-BmHsp12.6 using homologous DNA vaccine regimen or a heterologous prime boost approach. Control mice were immunized with vector alone or adjuvant. Isotype specific ELISA was performed as described in the methods section. Bars represent mean O.D ± SD from five mice per group. * Significant (p
    Figure Legend Snippet: Isotype of anti-BmHsp12.6 IgG antibodies in the sera of mice. Mice were immunized with A ) BmHsp12.6, B ) BmHsp12.6αc or C ) N-BmHsp12.6 using homologous DNA vaccine regimen or a heterologous prime boost approach. Control mice were immunized with vector alone or adjuvant. Isotype specific ELISA was performed as described in the methods section. Bars represent mean O.D ± SD from five mice per group. * Significant (p

    Techniques Used: Mouse Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

    Titer of IgG antibodies. Mice were immunized with A ) BmHsp12.6, B ) BmHsp12.6αc or C ) N-BmHsp12.6 using homologous DNA vaccine regimen or a heterologous prime boost approach. Approximately, 100 ng of peptides or proteins respectively (100 ng/100 µl/well) were coated on to the wells of an ELISA plate and bound serum IgG was detected using an HRP-labeled anti-mouse IgG secondary antibody. Each data point indicates mean (± S.D) value from five animals.
    Figure Legend Snippet: Titer of IgG antibodies. Mice were immunized with A ) BmHsp12.6, B ) BmHsp12.6αc or C ) N-BmHsp12.6 using homologous DNA vaccine regimen or a heterologous prime boost approach. Approximately, 100 ng of peptides or proteins respectively (100 ng/100 µl/well) were coated on to the wells of an ELISA plate and bound serum IgG was detected using an HRP-labeled anti-mouse IgG secondary antibody. Each data point indicates mean (± S.D) value from five animals.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Labeling

    Anti-rBmHsp12.6 IgG antibody isotypes in the human sera. IgG isotype antibodies against ( A ) rBmHsp12.6 protein, ( B ) BmHsp12.6αc peptide or ( C ) N-BmHsp12.6 in the sera of EN, MF, CP and NEN subjects were measured using an indirect ELISA. A total of 20 sera samples were evaluated from EN, MF, and CP from NEN. Each data point represents sera sample from a single individual. Each bar represents the mean ± SD of five patients from each group. * Significant (p
    Figure Legend Snippet: Anti-rBmHsp12.6 IgG antibody isotypes in the human sera. IgG isotype antibodies against ( A ) rBmHsp12.6 protein, ( B ) BmHsp12.6αc peptide or ( C ) N-BmHsp12.6 in the sera of EN, MF, CP and NEN subjects were measured using an indirect ELISA. A total of 20 sera samples were evaluated from EN, MF, and CP from NEN. Each data point represents sera sample from a single individual. Each bar represents the mean ± SD of five patients from each group. * Significant (p

    Techniques Used: Indirect ELISA

    Anti-rBmHsp12.6 IgG antibody levels in the sera of human. Levels of total IgG antibodies against ( A ) rBmHsp12.6 protein, ( B ) BmHsp12.6αc peptide or ( C ) N-BmHsp12.6 peptide in the sera of EN, MF, CP and NEN subjects were measured using an indirect ELISA. A total of 20 sera samples were evaluated from EN, MF, and CP and 10 samples from NEN. Each data point represents sera sample from a single individual. Horizontal lines represent geometric mean value. Data is represented as scatter plot where each dot represents absorbance of individual sera.
    Figure Legend Snippet: Anti-rBmHsp12.6 IgG antibody levels in the sera of human. Levels of total IgG antibodies against ( A ) rBmHsp12.6 protein, ( B ) BmHsp12.6αc peptide or ( C ) N-BmHsp12.6 peptide in the sera of EN, MF, CP and NEN subjects were measured using an indirect ELISA. A total of 20 sera samples were evaluated from EN, MF, and CP and 10 samples from NEN. Each data point represents sera sample from a single individual. Horizontal lines represent geometric mean value. Data is represented as scatter plot where each dot represents absorbance of individual sera.

    Techniques Used: Indirect ELISA

    16) Product Images from "Novel monoclonal antibody L2A5 specifically targeting sialyl-Tn and short glycans terminated by alpha-2–6 sialic acids"

    Article Title: Novel monoclonal antibody L2A5 specifically targeting sialyl-Tn and short glycans terminated by alpha-2–6 sialic acids

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-30421-w

    L2A5 isotype determination using capture ELISA. 1:1000 dilution of L2A5 mAb was used in a capture ELISA with IgG1, IgG2a, IgGb, IgG3, IgM and IgA. Results are expressed as mean of absorbance (Abs) at 450 nm + standard deviation. Antibodies with known isotype B72.3 (IgG1) and CD15S (IgM) were used as controls (CTR).
    Figure Legend Snippet: L2A5 isotype determination using capture ELISA. 1:1000 dilution of L2A5 mAb was used in a capture ELISA with IgG1, IgG2a, IgGb, IgG3, IgM and IgA. Results are expressed as mean of absorbance (Abs) at 450 nm + standard deviation. Antibodies with known isotype B72.3 (IgG1) and CD15S (IgM) were used as controls (CTR).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Standard Deviation

    Western blot analysis of anti-STn mAbs binding to STn-expressing cell lines and MUC1 STn – IgG. MDA-MB-231 STn + ( A ) and STn − ( B ) membrane extracts, as well as the chimeric protein MUC1 STn-IgG ( C .
    Figure Legend Snippet: Western blot analysis of anti-STn mAbs binding to STn-expressing cell lines and MUC1 STn – IgG. MDA-MB-231 STn + ( A ) and STn − ( B ) membrane extracts, as well as the chimeric protein MUC1 STn-IgG ( C .

    Techniques Used: Western Blot, Binding Assay, Expressing, Multiple Displacement Amplification

    17) Product Images from "Antibody responses to prime-boost vaccination with an HIV-1 gp145 Env protein and chimpanzee adenovirus vectors expressing HIV-1 gp140"

    Article Title: Antibody responses to prime-boost vaccination with an HIV-1 gp145 Env protein and chimpanzee adenovirus vectors expressing HIV-1 gp140

    Journal: AIDS (London, England)

    doi: 10.1097/QAD.0000000000001224

    Avidity of Env-specific Ab binding NaSCN-displacement ELISA performed (A) following last immunization: protein/AdC7 groups at wk 18, AdC7/protein groups at wk 20 and AdC7/AdC6 groups at wk 12 and (B) on week 12, 18, 20 for AdC7/protein 10 10 and AdC7/protein 10 9 to determine effect of additional protein boosts on avidity. Results are reported as the concentration of NaSCN (mean ± SEM) at which IgG binding was reduced by 50% as compared to binding in the absence of NaSCN. (A) Avidity of AdC7/protein 10 10 is significantly greater than both protein/AdC7 10 10 and AdC7/AdC6 10 10 , p =0.0009 and p =0.0439, respectively, while AdC7/AdC6 10 10 is greater than protein/AdC7 10 10 , p =0.0339. At the 10 9 dose, AdC7/protein and AdC7/AdC6 avidities are significantly greater than that of protein/AdC7, p =0.0030 and p =0.0164, respectively. (B) No significant change in avidity is seen in the AdC7/protein groups with administration of additional protein boosts. (all statistics performed by one-way ANOVA with Holm-Sidak multiple comparisons test).
    Figure Legend Snippet: Avidity of Env-specific Ab binding NaSCN-displacement ELISA performed (A) following last immunization: protein/AdC7 groups at wk 18, AdC7/protein groups at wk 20 and AdC7/AdC6 groups at wk 12 and (B) on week 12, 18, 20 for AdC7/protein 10 10 and AdC7/protein 10 9 to determine effect of additional protein boosts on avidity. Results are reported as the concentration of NaSCN (mean ± SEM) at which IgG binding was reduced by 50% as compared to binding in the absence of NaSCN. (A) Avidity of AdC7/protein 10 10 is significantly greater than both protein/AdC7 10 10 and AdC7/AdC6 10 10 , p =0.0009 and p =0.0439, respectively, while AdC7/AdC6 10 10 is greater than protein/AdC7 10 10 , p =0.0339. At the 10 9 dose, AdC7/protein and AdC7/AdC6 avidities are significantly greater than that of protein/AdC7, p =0.0030 and p =0.0164, respectively. (B) No significant change in avidity is seen in the AdC7/protein groups with administration of additional protein boosts. (all statistics performed by one-way ANOVA with Holm-Sidak multiple comparisons test).

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

    Ab responses are primarily IgG2 Isotyping was performed by ELISA following the last immunization: protein/AdC7 groups at wk 18, AdC7/protein groups at wk 20 and AdC7/AdC6 groups at wk 12. (A) At both AdC doses, AdC7/protein elicits a greater combined isotype response than protein/AdC7 and AdC7/AdC6 (10 10 dose p =0.0013, p =0.0034, respectively; 10 9 dose p =0.0085, p =0.0442, respectively). (B) Relative isotype frequency. Priming with protein versus AdC7 elicits a different isotype response independent of dose. Protein/AdC7 elicits predomoinantly IgG1 at a significantly greater frequency than both AdC7/protein and AdC7/AdC6, p
    Figure Legend Snippet: Ab responses are primarily IgG2 Isotyping was performed by ELISA following the last immunization: protein/AdC7 groups at wk 18, AdC7/protein groups at wk 20 and AdC7/AdC6 groups at wk 12. (A) At both AdC doses, AdC7/protein elicits a greater combined isotype response than protein/AdC7 and AdC7/AdC6 (10 10 dose p =0.0013, p =0.0034, respectively; 10 9 dose p =0.0085, p =0.0442, respectively). (B) Relative isotype frequency. Priming with protein versus AdC7 elicits a different isotype response independent of dose. Protein/AdC7 elicits predomoinantly IgG1 at a significantly greater frequency than both AdC7/protein and AdC7/AdC6, p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    18) Product Images from "Antibodies to the GABAB receptor in limbic encephalitis with seizures: case series and characterisation of the antigen"

    Article Title: Antibodies to the GABAB receptor in limbic encephalitis with seizures: case series and characterisation of the antigen

    Journal: Lancet neurology

    doi: 10.1016/S1474-4422(09)70324-2

    Culture of rat hippocampal neurons incubated (live, non-permeabilised) with the CSF of a patient with limbic encephalitis and a control individual Note the intense punctate reactivity of patient's antibodies with cell surface antigens (A) and the absence of reactivity in the control (B); nuclei of neurons stained with 4',6-diamidino-2-phenylindole (DAPI). The surface antigens were precipitated using the antibodies within the patient's serum, and then electrophoretically separated and visualised with EZBlue (C). Patient's antibodies (P) precipitated two main protein bands at about 105 kDa and 90 kDa; these bands are not seen in the precipitate using serum from a control individual (N). Sequencing of the 105 kDa band by use of mass spectrometry showed it contained the B1 and B2 subunits of the GABA B ). The 90 kDa and other smaller bands were proteolytic fragments and patient's IgG products. Subsequent transfer of the gel to nitrocellulose and immunoblotting with antibodies specific for each of the GABA B (D) subunits confirmed that patient's antibodies precipitated the B1 and B2 subunits (105 kDa) and that the 90 kDa band was a proteolytic fragment of B1.
    Figure Legend Snippet: Culture of rat hippocampal neurons incubated (live, non-permeabilised) with the CSF of a patient with limbic encephalitis and a control individual Note the intense punctate reactivity of patient's antibodies with cell surface antigens (A) and the absence of reactivity in the control (B); nuclei of neurons stained with 4',6-diamidino-2-phenylindole (DAPI). The surface antigens were precipitated using the antibodies within the patient's serum, and then electrophoretically separated and visualised with EZBlue (C). Patient's antibodies (P) precipitated two main protein bands at about 105 kDa and 90 kDa; these bands are not seen in the precipitate using serum from a control individual (N). Sequencing of the 105 kDa band by use of mass spectrometry showed it contained the B1 and B2 subunits of the GABA B ). The 90 kDa and other smaller bands were proteolytic fragments and patient's IgG products. Subsequent transfer of the gel to nitrocellulose and immunoblotting with antibodies specific for each of the GABA B (D) subunits confirmed that patient's antibodies precipitated the B1 and B2 subunits (105 kDa) and that the 90 kDa band was a proteolytic fragment of B1.

    Techniques Used: Incubation, Staining, Sequencing, Mass Spectrometry

    19) Product Images from "Evaluation of Immunogenicity of Novel Isoform of EG95 (EG95-5G1) From Echinococcus granulosus in BALB/C Mice"

    Article Title: Evaluation of Immunogenicity of Novel Isoform of EG95 (EG95-5G1) From Echinococcus granulosus in BALB/C Mice

    Journal: Iranian Journal of Parasitology

    doi:

    Results for immunoglubin G subtypes evaluation IgG subtypes in mice serum samples of different experimental groups were assayed against rEG95 by ELISA. Data from each group was compared by other groups with LSD test via one-way analysis of variance (one-way ANOVA). P -values less than 0.05 were recognized as significant. Freund’s : sera from mice immunized with rEG95 formulated with FA. Alum : sera from mice immunized with rEG95 formulated with alum. Trx : sera from mice immunized with Trx pET32 *: significant difference with non-immunized (negative control) group ( P
    Figure Legend Snippet: Results for immunoglubin G subtypes evaluation IgG subtypes in mice serum samples of different experimental groups were assayed against rEG95 by ELISA. Data from each group was compared by other groups with LSD test via one-way analysis of variance (one-way ANOVA). P -values less than 0.05 were recognized as significant. Freund’s : sera from mice immunized with rEG95 formulated with FA. Alum : sera from mice immunized with rEG95 formulated with alum. Trx : sera from mice immunized with Trx pET32 *: significant difference with non-immunized (negative control) group ( P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Negative Control

    20) Product Images from "Identification of a phage-encoded immunoglobulin-binding protein from invasive Neisseria meningitidis"

    Article Title: Identification of a phage-encoded immunoglobulin-binding protein from invasive Neisseria meningitidis

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1301153

    Isolation of Nm proteins binding to human IgG by precipitation with Protein A or G beads. NmW135 strain A22 was cultured in CDM+HuS or MH as indicated. Unmarked lanes were intentionally left unloaded. The arrows indicate bands that produced a common set
    Figure Legend Snippet: Isolation of Nm proteins binding to human IgG by precipitation with Protein A or G beads. NmW135 strain A22 was cultured in CDM+HuS or MH as indicated. Unmarked lanes were intentionally left unloaded. The arrows indicate bands that produced a common set

    Techniques Used: Isolation, Binding Assay, Cell Culture, Produced

    Immunoglobulin-binding protein activity exhibited by diverse Nm strains. Panel a. shows IgG binding to diverse Nm strains with serum from two donors. Panel b. shows changes in IgG binding to Nm strains after each of three rounds of serial growth (indicated
    Figure Legend Snippet: Immunoglobulin-binding protein activity exhibited by diverse Nm strains. Panel a. shows IgG binding to diverse Nm strains with serum from two donors. Panel b. shows changes in IgG binding to Nm strains after each of three rounds of serial growth (indicated

    Techniques Used: Binding Assay, Activity Assay

    Immunoglobulin-binding activity of recombinant TspB derivatives by ELISA. Panel a. shows IgG-binding activity in whole serum from donor 2 of full length (FL) TspB1628 (filled circles), TspB1628 CRPro (filled squares), TspB1628 CR (filled triangles), TspB1548
    Figure Legend Snippet: Immunoglobulin-binding activity of recombinant TspB derivatives by ELISA. Panel a. shows IgG-binding activity in whole serum from donor 2 of full length (FL) TspB1628 (filled circles), TspB1628 CRPro (filled squares), TspB1628 CR (filled triangles), TspB1548

    Techniques Used: Binding Assay, Activity Assay, Recombinant, Enzyme-linked Immunosorbent Assay

    Effect of bacterial culture conditions on immunoglobulin-binding activity of NmW135 strain A22 and NmB strain H44/76 by flow cytometry. Bacteria were cultured in the indicated media. The rightward shift to higher fluorescence indicates bound human IgG
    Figure Legend Snippet: Effect of bacterial culture conditions on immunoglobulin-binding activity of NmW135 strain A22 and NmB strain H44/76 by flow cytometry. Bacteria were cultured in the indicated media. The rightward shift to higher fluorescence indicates bound human IgG

    Techniques Used: Binding Assay, Activity Assay, Flow Cytometry, Cytometry, Cell Culture, Fluorescence

    Biofilm containing TspB, IgG, and DNA enveloping aggregates of NmB cultured in the presence of human serum. Laser scanning confocal micrographs (63x) of an aggregate of NmB strain H44/76 cultured in CDM+HuS and mouse anti-TspB1628 CR. Human IgG is marked
    Figure Legend Snippet: Biofilm containing TspB, IgG, and DNA enveloping aggregates of NmB cultured in the presence of human serum. Laser scanning confocal micrographs (63x) of an aggregate of NmB strain H44/76 cultured in CDM+HuS and mouse anti-TspB1628 CR. Human IgG is marked

    Techniques Used: Cell Culture

    21) Product Images from "Identification of two liver proteins recognized by autoantibodies elicited in mice infected with mouse hepatitis virus A59"

    Article Title: Identification of two liver proteins recognized by autoantibodies elicited in mice infected with mouse hepatitis virus A59

    Journal: European Journal of Immunology

    doi: 10.1002/1521-4141(200105)31:5 < 1447::AID-IMMU1447 > 3.0.CO;2-6

    Reactivity of sera from MHV‐infected CBA/Ht mice with extracts of several tissues from non‐infected CBA/Ht mice (A) or liver lysate from different sources (B). Organ lysates were prepared as indicated in Sect. 4 and run in SDS‐PAGE using 10% gels, transferred onto nitrocellulose sheets and incubated with a 1:100 serum dilution. Bound antibodies were revealed with peroxidase‐labeled IgG anti‐mouse IgG and ECL reagents. Results were obtained with pooled serum from four mice. Control serum (–) means serum obtained before infection and (+) indicates serum corresponding to 8 weeks post‐infection with MHV. The positions of molecular mass markers (kDa) are shown at right. WILEY‐VCH
    Figure Legend Snippet: Reactivity of sera from MHV‐infected CBA/Ht mice with extracts of several tissues from non‐infected CBA/Ht mice (A) or liver lysate from different sources (B). Organ lysates were prepared as indicated in Sect. 4 and run in SDS‐PAGE using 10% gels, transferred onto nitrocellulose sheets and incubated with a 1:100 serum dilution. Bound antibodies were revealed with peroxidase‐labeled IgG anti‐mouse IgG and ECL reagents. Results were obtained with pooled serum from four mice. Control serum (–) means serum obtained before infection and (+) indicates serum corresponding to 8 weeks post‐infection with MHV. The positions of molecular mass markers (kDa) are shown at right. WILEY‐VCH

    Techniques Used: Infection, Crocin Bleaching Assay, Mouse Assay, SDS Page, Incubation, Labeling

    Time course of Ab to the 40‐kDa liver protein and plasma Ig level in MHV‐infected CBA/Ht mice. Upper panel: Western‐blot reactivity of serum from MHV‐infected CBA/Ht with the 40‐kDa liver protein (see Fig. 1 for details); (+) means that serum from all animals tested reacted with the mouse liver 40‐kDa protein; (–) means absence of reactivity. Lower panel: Total IgG concentration of the corresponding sera was determined as indicated in Sect. 4. Values correspond to individual mice. Results were subject to statistical analysis by using the Mann‐Whitney U‐test. * p
    Figure Legend Snippet: Time course of Ab to the 40‐kDa liver protein and plasma Ig level in MHV‐infected CBA/Ht mice. Upper panel: Western‐blot reactivity of serum from MHV‐infected CBA/Ht with the 40‐kDa liver protein (see Fig. 1 for details); (+) means that serum from all animals tested reacted with the mouse liver 40‐kDa protein; (–) means absence of reactivity. Lower panel: Total IgG concentration of the corresponding sera was determined as indicated in Sect. 4. Values correspond to individual mice. Results were subject to statistical analysis by using the Mann‐Whitney U‐test. * p

    Techniques Used: Infection, Crocin Bleaching Assay, Mouse Assay, Western Blot, Concentration Assay, MANN-WHITNEY

    22) Product Images from "Analysis of Pre-existing IgG and IgM Antibodies against Polyethylene Glycol (PEG) in the General Population"

    Article Title: Analysis of Pre-existing IgG and IgM Antibodies against Polyethylene Glycol (PEG) in the General Population

    Journal: Analytical chemistry

    doi: 10.1021/acs.analchem.6b03437

    Anti-PEG IgM, IgG, IgG1, and IgG2 prevalence by (A–D) age group, (E–H) gender, and (I–L) race in healthy individuals ( n = 377).
    Figure Legend Snippet: Anti-PEG IgM, IgG, IgG1, and IgG2 prevalence by (A–D) age group, (E–H) gender, and (I–L) race in healthy individuals ( n = 377).

    Techniques Used:

    Anti-PEG IgM, IgG, IgG1, and IgG2 levels by (A–D) age group, (E–H) gender, and (I–L) race in healthy individuals ( n = 377). The data are depicted using Tukey’s method for box-and-whisker plots, with samples outside of the whiskers shown as open circles.
    Figure Legend Snippet: Anti-PEG IgM, IgG, IgG1, and IgG2 levels by (A–D) age group, (E–H) gender, and (I–L) race in healthy individuals ( n = 377). The data are depicted using Tukey’s method for box-and-whisker plots, with samples outside of the whiskers shown as open circles.

    Techniques Used: Whisker Assay

    23) Product Images from "Partial Immunity to Fasciola hepatica in Mice after Vaccination with FhSAP2 Delivered as Recombinant Protein or DNA Construct"

    Article Title: Partial Immunity to Fasciola hepatica in Mice after Vaccination with FhSAP2 Delivered as Recombinant Protein or DNA Construct

    Journal: Ethnicity & disease

    doi:

    (A) Levels of specific anti-FhSAP2 IgG 1 , IgG 2a , IgG 2b and IgG 3 elicited by immunization with FhSAP2, or with cDNA-FhSAP2 45 days after challenge. Values represent the mean ± SD of triplicate ELISA readings of each vaccinated group. PC represent
    Figure Legend Snippet: (A) Levels of specific anti-FhSAP2 IgG 1 , IgG 2a , IgG 2b and IgG 3 elicited by immunization with FhSAP2, or with cDNA-FhSAP2 45 days after challenge. Values represent the mean ± SD of triplicate ELISA readings of each vaccinated group. PC represent

    Techniques Used: Enzyme-linked Immunosorbent Assay

    24) Product Images from "Plasmodium vivax Cell-Traversal Protein for Ookinetes and Sporozoites: Naturally Acquired Humoral Immune Response and B-Cell Epitope Mapping in Brazilian Amazon Inhabitants"

    Article Title: Plasmodium vivax Cell-Traversal Protein for Ookinetes and Sporozoites: Naturally Acquired Humoral Immune Response and B-Cell Epitope Mapping in Brazilian Amazon Inhabitants

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.00077

    Comparison of profiles of high responders (HRs), low responders (LRs), and non-responders (NRs) to PvCelTOS . (A) Diagram of epidemiological data. The mean values of epidemiological parameters of HRs, LRs, and NRs to PvCelTOS are represented by red, blue, and yellow areas, respectively. Age and time of residence in endemic areas are expressed in years, while time elapsed from the last malaria episode is expressed in months and frequency of recent malaria episodes indicates the percentage of individuals who reported malaria episodes in the last year. HRs presented a higher number of previous malaria episodes and higher frequency of recent malaria episodes than LRs ( p = 0.0051 and p = 0.02, respectively) (B) Comparison of ratio (cytophilic/non-cytophilic) antibodies. Red and blue points represent the HR and LR, respectively. Points above the value of 1 represent individuals with a cytophilic profile of IgG (IgG1 + IgG3, higher than IgG2 + IgG4).
    Figure Legend Snippet: Comparison of profiles of high responders (HRs), low responders (LRs), and non-responders (NRs) to PvCelTOS . (A) Diagram of epidemiological data. The mean values of epidemiological parameters of HRs, LRs, and NRs to PvCelTOS are represented by red, blue, and yellow areas, respectively. Age and time of residence in endemic areas are expressed in years, while time elapsed from the last malaria episode is expressed in months and frequency of recent malaria episodes indicates the percentage of individuals who reported malaria episodes in the last year. HRs presented a higher number of previous malaria episodes and higher frequency of recent malaria episodes than LRs ( p = 0.0051 and p = 0.02, respectively) (B) Comparison of ratio (cytophilic/non-cytophilic) antibodies. Red and blue points represent the HR and LR, respectively. Points above the value of 1 represent individuals with a cytophilic profile of IgG (IgG1 + IgG3, higher than IgG2 + IgG4).

    Techniques Used:

    Reactivity index (RI) of IgG and subclass against PvCelTOS . On both graphs, each point represents an individual RI against PvCelTOS and the red traced line represents the cutoff. Ninety-four individuals presented RI against PvCelTOS higher than 1 and were considered responders to this protein. Among the responders, IgG1 was the prevalent subclass in comparison to IgG2 ( p = 0.0012), IgG3 ( p
    Figure Legend Snippet: Reactivity index (RI) of IgG and subclass against PvCelTOS . On both graphs, each point represents an individual RI against PvCelTOS and the red traced line represents the cutoff. Ninety-four individuals presented RI against PvCelTOS higher than 1 and were considered responders to this protein. Among the responders, IgG1 was the prevalent subclass in comparison to IgG2 ( p = 0.0012), IgG3 ( p

    Techniques Used:

    Mapping of B-cell epitopes in PvCelTOS . Each column represents a peptide, the numbers indicate the first and last amino acid (aa-aa) of the peptide. The points represent the value of IgG reactivity index (RI) specific for each peptide of one responder to PvCelTOS and the red traced line represents the cutoff value. The black lines indicate median and interquartile range. If the RI for one peptide was higher than 1, the individual was considered positive to this peptide. The white bar on top represents the linear structure of the protein, in which the blue boxes indicate the BepiPred prediction score and red boxes indicate the Emini surface accessibility score of predicted linear epitopes.
    Figure Legend Snippet: Mapping of B-cell epitopes in PvCelTOS . Each column represents a peptide, the numbers indicate the first and last amino acid (aa-aa) of the peptide. The points represent the value of IgG reactivity index (RI) specific for each peptide of one responder to PvCelTOS and the red traced line represents the cutoff value. The black lines indicate median and interquartile range. If the RI for one peptide was higher than 1, the individual was considered positive to this peptide. The white bar on top represents the linear structure of the protein, in which the blue boxes indicate the BepiPred prediction score and red boxes indicate the Emini surface accessibility score of predicted linear epitopes.

    Techniques Used:

    25) Product Images from "A Role for the Transcription Factor Arid3a in Mouse B2 Lymphocyte Expansion and Peritoneal B1a Generation"

    Article Title: A Role for the Transcription Factor Arid3a in Mouse B2 Lymphocyte Expansion and Peritoneal B1a Generation

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.01387

    A requirement for Arid3a in humoral immunity. (A) Plasma was collected from naïve mice of the indicated genotypes aged 10–12 weeks old and analyzed for total IgM, IgA, and IgG levels with Enzyme-Linked ImmunoSorbent Assay. (B) IgG1, IgG2b, and IgG3 antibody titers. n = 15–26 for each group and p -values were determined by Student’s t -test.
    Figure Legend Snippet: A requirement for Arid3a in humoral immunity. (A) Plasma was collected from naïve mice of the indicated genotypes aged 10–12 weeks old and analyzed for total IgM, IgA, and IgG levels with Enzyme-Linked ImmunoSorbent Assay. (B) IgG1, IgG2b, and IgG3 antibody titers. n = 15–26 for each group and p -values were determined by Student’s t -test.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    26) Product Images from "Dobrava, but not Saaremaa, hantavirus is lethal and induces nitric oxide production in suckling mice"

    Article Title: Dobrava, but not Saaremaa, hantavirus is lethal and induces nitric oxide production in suckling mice

    Journal: Microbes and Infection

    doi: 10.1016/j.micinf.2005.09.010

    Antibody responses in C57/BL6 and iNOS−/− mice after HTNV infection. (A) IgG subclass specific response against N protein. (B) Neutralizing titers against HTNV. The data represent mean + SD.
    Figure Legend Snippet: Antibody responses in C57/BL6 and iNOS−/− mice after HTNV infection. (A) IgG subclass specific response against N protein. (B) Neutralizing titers against HTNV. The data represent mean + SD.

    Techniques Used: Mouse Assay, Infection

    IgG2a/IgG1 ratio in intraperitoneally infected BALB/c mice 43 days after infection. Data shown represent mean ± SD of six mice/group.
    Figure Legend Snippet: IgG2a/IgG1 ratio in intraperitoneally infected BALB/c mice 43 days after infection. Data shown represent mean ± SD of six mice/group.

    Techniques Used: Infection, Mouse Assay

    Total IgG and IgG subclass responses against N in sera from adult NMRI mice 60 days after DOBV, HTNV, PUUV Kazan-wt and PUUV Kazan-E6 p6 inoculation and 49 days after SAAV inoculation. (A) IgG anti-N responses. (B) IgG subclass anti-N responses in pooled sera from four mice inoculated intravenously with PUUV Kazan-wt (1000 bank vole ID 50 ), four mice inoculated intravenously with PUUV Kazan-E6 (25,000 FFU), three mice inoculated intravenously with HTNV (90,000 FFU), four mice inoculated intravenously and subcutaneously with DOBV (80,000 FFU), and eight mice inoculated intravenously and subcutaneously with SAAV (100,000 FFU).
    Figure Legend Snippet: Total IgG and IgG subclass responses against N in sera from adult NMRI mice 60 days after DOBV, HTNV, PUUV Kazan-wt and PUUV Kazan-E6 p6 inoculation and 49 days after SAAV inoculation. (A) IgG anti-N responses. (B) IgG subclass anti-N responses in pooled sera from four mice inoculated intravenously with PUUV Kazan-wt (1000 bank vole ID 50 ), four mice inoculated intravenously with PUUV Kazan-E6 (25,000 FFU), three mice inoculated intravenously with HTNV (90,000 FFU), four mice inoculated intravenously and subcutaneously with DOBV (80,000 FFU), and eight mice inoculated intravenously and subcutaneously with SAAV (100,000 FFU).

    Techniques Used: Mouse Assay

    27) Product Images from "Recurrent Membranous Nephropathy in an Allograft Caused by IgG3? Targeting the PLA2 Receptor"

    Article Title: Recurrent Membranous Nephropathy in an Allograft Caused by IgG3? Targeting the PLA2 Receptor

    Journal: Journal of the American Society of Nephrology : JASN

    doi: 10.1681/ASN.2012060577

    Characterization of immune deposits in kidney biopsy specimens from grafted (A–D) and native (E) kidneys. (A) Immunofluorescence study showing early recurrence of the MN (day 13) characterized by granular deposits of IgG. (B) Representative segment of the capillary wall analyzed by electron microscopy. Electron-dense deposits seen on the outer aspect of the glomerular basement membrane do not show any organization. (C) Immunostaining for IgG subclasses and light-chain isotypes showing the presence of monotypic IgG3κ. (D) Complement components, including C3, C1q and C5b-9, detected in the absence of MBL. (E) Kidney biopsy specimen from native kidney stained for IgG3 and light-chain isotype. The specimens shown in E are paraffin sections, whereas those shown in A, C, and D are cryostat sections. Original magnification for A, C, D, and E x400; for B x40,000.
    Figure Legend Snippet: Characterization of immune deposits in kidney biopsy specimens from grafted (A–D) and native (E) kidneys. (A) Immunofluorescence study showing early recurrence of the MN (day 13) characterized by granular deposits of IgG. (B) Representative segment of the capillary wall analyzed by electron microscopy. Electron-dense deposits seen on the outer aspect of the glomerular basement membrane do not show any organization. (C) Immunostaining for IgG subclasses and light-chain isotypes showing the presence of monotypic IgG3κ. (D) Complement components, including C3, C1q and C5b-9, detected in the absence of MBL. (E) Kidney biopsy specimen from native kidney stained for IgG3 and light-chain isotype. The specimens shown in E are paraffin sections, whereas those shown in A, C, and D are cryostat sections. Original magnification for A, C, D, and E x400; for B x40,000.

    Techniques Used: Immunofluorescence, Electron Microscopy, Immunostaining, Staining

    Detection of PLA2R in glomerular immune deposits. Immunofluorescence analysis of paraffin kidney biopsy specimens shows PLA2R in (A) native and (B) grafted kidney. (C–E) Positively stained glomeruli in a biopsy specimen from grafted kidney that has been double-labeled with anti-PLA2R (C) and anti-human IgG3 antibodies (D); E shows the merged image of boxed areas in C and D. (F) Quantitative analysis of the fluorescence recorded across sections of a representative capillary loop (arrowheads). Note superimposition of the two signals, which indicates that subepithelial immune deposits are composed of PLA2R (red) and IgG3 (green). Original magnification for A, B, C, D, and E x400.
    Figure Legend Snippet: Detection of PLA2R in glomerular immune deposits. Immunofluorescence analysis of paraffin kidney biopsy specimens shows PLA2R in (A) native and (B) grafted kidney. (C–E) Positively stained glomeruli in a biopsy specimen from grafted kidney that has been double-labeled with anti-PLA2R (C) and anti-human IgG3 antibodies (D); E shows the merged image of boxed areas in C and D. (F) Quantitative analysis of the fluorescence recorded across sections of a representative capillary loop (arrowheads). Note superimposition of the two signals, which indicates that subepithelial immune deposits are composed of PLA2R (red) and IgG3 (green). Original magnification for A, B, C, D, and E x400.

    Techniques Used: Immunofluorescence, Staining, Labeling, Fluorescence

    28) Product Images from "Evaluation of Wuchereria bancrofti GST as a Vaccine Candidate for Lymphatic Filariasis"

    Article Title: Evaluation of Wuchereria bancrofti GST as a Vaccine Candidate for Lymphatic Filariasis

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0000457

    Subclass specific anti-WbGST IgG antibodies in the sera of mice immunized with rWbGST protein. Subclass analysis was performed using a mouse antibody isotyping ELISA kit. The bars represent the mean O.D.±SD at 405 nm of five mice per group.
    Figure Legend Snippet: Subclass specific anti-WbGST IgG antibodies in the sera of mice immunized with rWbGST protein. Subclass analysis was performed using a mouse antibody isotyping ELISA kit. The bars represent the mean O.D.±SD at 405 nm of five mice per group.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Human immune responses to WbGST. A. Immunoreactivity of rWbGST with sera from different clinical groups of bancroftian filariasis. Approximately 1 µg of rWbGST was resolved on 15% SDS-PAGE, transferred to nitrocellulose membrane and blots were probed with pooled sera from MF, CP, EN or NEN individuals. Results showed strong immunoreactivity with pooled EN sera followed by CP and MF but no reactivity was detected with control NEN sera. Data is representative of one of three experiments using the same sera samples. B. Humoral immune response to rWbGST in human subjects. Total IgG levels against rWbGST in various clinical groups were evaluated by ELISA. Each data point represents single individual absorbance from the four different groups. Horizontal lines represent geometric mean value of EN (43), Mf (45), CP (45) and NEN (39) samples respectively. C. WbGST-specific IgG subclasses in the sera from different clinical groups. Isotype of anti-WbGST IgG antibodies in the sera from various groups of human subjects (EN, MF, CP, TPE and NEN) were evaluated using an isotype-specific ELISA. Data presented is mean±SD value from EN (43), Mf (45), CP (45) and NEN (39). * Significant (p
    Figure Legend Snippet: Human immune responses to WbGST. A. Immunoreactivity of rWbGST with sera from different clinical groups of bancroftian filariasis. Approximately 1 µg of rWbGST was resolved on 15% SDS-PAGE, transferred to nitrocellulose membrane and blots were probed with pooled sera from MF, CP, EN or NEN individuals. Results showed strong immunoreactivity with pooled EN sera followed by CP and MF but no reactivity was detected with control NEN sera. Data is representative of one of three experiments using the same sera samples. B. Humoral immune response to rWbGST in human subjects. Total IgG levels against rWbGST in various clinical groups were evaluated by ELISA. Each data point represents single individual absorbance from the four different groups. Horizontal lines represent geometric mean value of EN (43), Mf (45), CP (45) and NEN (39) samples respectively. C. WbGST-specific IgG subclasses in the sera from different clinical groups. Isotype of anti-WbGST IgG antibodies in the sera from various groups of human subjects (EN, MF, CP, TPE and NEN) were evaluated using an isotype-specific ELISA. Data presented is mean±SD value from EN (43), Mf (45), CP (45) and NEN (39). * Significant (p

    Techniques Used: SDS Page, Enzyme-linked Immunosorbent Assay

    29) Product Images from "Protection against Candidiasis by an Immunoglobulin G3 (IgG3) Monoclonal Antibody Specific for the Same Mannotriose as an IgM Protective Antibody"

    Article Title: Protection against Candidiasis by an Immunoglobulin G3 (IgG3) Monoclonal Antibody Specific for the Same Mannotriose as an IgM Protective Antibody

    Journal: Infection and Immunity

    doi:

    The C3.1 epitope is uniformly distributed on the cell surface of yeast forms of C. albicans . Hydrophilic stationary-phase yeast cells were reacted with MAb C3.1, washed, and counterreacted with fluorescence-labeled anti-mouse IgG. The C3.1 epitope is located over the entire cell surface (A). The same field was photographed under phase-contrast microscopy (B). The C3.1 epitope distribution displays the same homogeneous pattern over the cell surface as the epitope recognized by the IgM protective antibody, MAb IgM B6.1. These results provide further evidence that the IgG3 antibody is specific for the same oligomannoside as the protective IgM. Note that some yeast cells (arrows) and all of the blastoconidia (arrowheads) were nonreactive with MAb C3.1. The importance of nonreactive cells in pathogenesis and protection experiments is unknown. Bar, 10 μM.
    Figure Legend Snippet: The C3.1 epitope is uniformly distributed on the cell surface of yeast forms of C. albicans . Hydrophilic stationary-phase yeast cells were reacted with MAb C3.1, washed, and counterreacted with fluorescence-labeled anti-mouse IgG. The C3.1 epitope is located over the entire cell surface (A). The same field was photographed under phase-contrast microscopy (B). The C3.1 epitope distribution displays the same homogeneous pattern over the cell surface as the epitope recognized by the IgM protective antibody, MAb IgM B6.1. These results provide further evidence that the IgG3 antibody is specific for the same oligomannoside as the protective IgM. Note that some yeast cells (arrows) and all of the blastoconidia (arrowheads) were nonreactive with MAb C3.1. The importance of nonreactive cells in pathogenesis and protection experiments is unknown. Bar, 10 μM.

    Techniques Used: Fluorescence, Labeling, Microscopy

    30) Product Images from "Evaluation of the acquired immune responses to Plasmodium vivax VIR variant antigens in individuals living in malaria-endemic areas of Brazil"

    Article Title: Evaluation of the acquired immune responses to Plasmodium vivax VIR variant antigens in individuals living in malaria-endemic areas of Brazil

    Journal: Malaria Journal

    doi: 10.1186/1475-2875-5-83

    Association between the percentage of responders that recognize each recombinant protein and the number of previous episodes malaria . A total of 186 individuals were grouped according to the number of times they had episodes of P. vivax malaria. The cut-off values were the same as used for Table 1. There was a significant increase in the percentage of individuals containing IgG anti-AMA-1 in the groups with previous malaria episodes when compared with individuals without previous episodes of the disease ( P
    Figure Legend Snippet: Association between the percentage of responders that recognize each recombinant protein and the number of previous episodes malaria . A total of 186 individuals were grouped according to the number of times they had episodes of P. vivax malaria. The cut-off values were the same as used for Table 1. There was a significant increase in the percentage of individuals containing IgG anti-AMA-1 in the groups with previous malaria episodes when compared with individuals without previous episodes of the disease ( P

    Techniques Used: Recombinant

    31) Product Images from "Immunoglobulin Class Switch Recombination Is Impaired in Atm-deficient Mice"

    Article Title: Immunoglobulin Class Switch Recombination Is Impaired in Atm-deficient Mice

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20041074

    Defective Ig production in Atm −/− B cells activated in vitro. CD19 + B cells were isolated from the spleens of Atm −/− and WT mice and cultured with CD40L and IL-4 (IgG1, IgG2a, and IgM), CD40L, IL-4, IL-5, TGF-β, and anti-IgD dextran (IgA) or LPS (IgG2b, IgG3). Results are displayed as the mean of three to four experiments ± SEM Atm −/− titer as a percentage of the Atm +/+ controls.
    Figure Legend Snippet: Defective Ig production in Atm −/− B cells activated in vitro. CD19 + B cells were isolated from the spleens of Atm −/− and WT mice and cultured with CD40L and IL-4 (IgG1, IgG2a, and IgM), CD40L, IL-4, IL-5, TGF-β, and anti-IgD dextran (IgA) or LPS (IgG2b, IgG3). Results are displayed as the mean of three to four experiments ± SEM Atm −/− titer as a percentage of the Atm +/+ controls.

    Techniques Used: In Vitro, Isolation, Mouse Assay, Cell Culture

    Real-time RT-PCR analysis of germline and productive IgG1 and IgE transcripts in Atm −/− B cells activated in vitro. Total RNA was isolated from B cells activated in vitro, and quantitative real-time RT-PCR was performed as described in Materials and Methods. Germline transcripts were measured at day 3 of culture and productive transcripts were measured at day 6 of culture. The data were normalized to GAPDH and presented as the increase in transcript level over unstimulated wild-type B cells. The data shown are representative of two independent experiments.
    Figure Legend Snippet: Real-time RT-PCR analysis of germline and productive IgG1 and IgE transcripts in Atm −/− B cells activated in vitro. Total RNA was isolated from B cells activated in vitro, and quantitative real-time RT-PCR was performed as described in Materials and Methods. Germline transcripts were measured at day 3 of culture and productive transcripts were measured at day 6 of culture. The data were normalized to GAPDH and presented as the increase in transcript level over unstimulated wild-type B cells. The data shown are representative of two independent experiments.

    Techniques Used: Quantitative RT-PCR, In Vitro, Isolation

    Surface Ig expression of IgG1 and IgE isotypes on in vitro–stimulated wild type and Atm −/− B cells. Cells were harvested after 4 d of in vitro stimulation with CD40L and IL-4, and flow cytometric analysis was used to determine surface IgG1 and IgE expression. Numbers on the dot plots show the percentage of switched cells as a proportion of B220 + B cells. Results are representative of four independent experiments.
    Figure Legend Snippet: Surface Ig expression of IgG1 and IgE isotypes on in vitro–stimulated wild type and Atm −/− B cells. Cells were harvested after 4 d of in vitro stimulation with CD40L and IL-4, and flow cytometric analysis was used to determine surface IgG1 and IgE expression. Numbers on the dot plots show the percentage of switched cells as a proportion of B220 + B cells. Results are representative of four independent experiments.

    Techniques Used: Expressing, In Vitro, Flow Cytometry

    Clonal expansion of IgG1-switched B cells. CFSE-stained resting B cells were stimulated in vitro with CD40L and IL-4 for 3–6 d and counterstained with goat anti–mouse IgG1 as described in Materials and Methods. (A) CFSE staining profiles of Atm +/+ and Atm −/− B cells are presented in the first two columns. The third column shows the percentage of recovered Atm −/− and Atm +/+ cells that have undergone the indicated number of cell divisions. (B) Two-color flow cytometric profiles indicate IgG1 expression on CFSE stained cells. The percentage of B cells expressing IgG1 is indicated for Atm +/+ B cells and Atm −/− B cells that have undergone the indicated numbers of cell divisions. Results are representative of three independent experiments.
    Figure Legend Snippet: Clonal expansion of IgG1-switched B cells. CFSE-stained resting B cells were stimulated in vitro with CD40L and IL-4 for 3–6 d and counterstained with goat anti–mouse IgG1 as described in Materials and Methods. (A) CFSE staining profiles of Atm +/+ and Atm −/− B cells are presented in the first two columns. The third column shows the percentage of recovered Atm −/− and Atm +/+ cells that have undergone the indicated number of cell divisions. (B) Two-color flow cytometric profiles indicate IgG1 expression on CFSE stained cells. The percentage of B cells expressing IgG1 is indicated for Atm +/+ B cells and Atm −/− B cells that have undergone the indicated numbers of cell divisions. Results are representative of three independent experiments.

    Techniques Used: Staining, In Vitro, Flow Cytometry, Expressing

    Impaired Ig production in Atm −/− mice. (A) Sera from eight Atm -deficient mice and littermate controls were assayed for total Ig levels by ELISA. IgE was undetectable by ELISA in sera from unimmunized mice. Results are displayed as the mean ± SEM Atm −/− titer as a percentage of the Atm +/+ controls. (B–D) Three Atm −/− mice and their WT littermates were immunized with TNP-KLH/Alum i.p., and serum was collected at the indicated time points. Serum levels of Ag-specific IgM (B) and IgG1 (C) were assayed by ELISA; data points represent the geometric mean ± geometric SEM of the ELISA titers. (D) Mice received a second immunization of TNP-KLH/Alum 40 d after the first immunization, and serum was collected after 8 d. Serum levels of total IgE and Ag-specific IgG2b and IgG3 were assayed by ELISA, and data points are displayed as the mean ± SD Atm −/− titer as a percentage of the Atm +/+ controls. The dotted line indicates the limit of detection. IgE, IgG2b, and IgG3 were undetectable in Atm −/− mice. Results are representative of two independent experiments. (E) Serum levels of Ag-specific IgA were measured in mice (five per group) immunized with TNP-KLH p.o., and serum was collected 4 d after the second immunization.
    Figure Legend Snippet: Impaired Ig production in Atm −/− mice. (A) Sera from eight Atm -deficient mice and littermate controls were assayed for total Ig levels by ELISA. IgE was undetectable by ELISA in sera from unimmunized mice. Results are displayed as the mean ± SEM Atm −/− titer as a percentage of the Atm +/+ controls. (B–D) Three Atm −/− mice and their WT littermates were immunized with TNP-KLH/Alum i.p., and serum was collected at the indicated time points. Serum levels of Ag-specific IgM (B) and IgG1 (C) were assayed by ELISA; data points represent the geometric mean ± geometric SEM of the ELISA titers. (D) Mice received a second immunization of TNP-KLH/Alum 40 d after the first immunization, and serum was collected after 8 d. Serum levels of total IgE and Ag-specific IgG2b and IgG3 were assayed by ELISA, and data points are displayed as the mean ± SD Atm −/− titer as a percentage of the Atm +/+ controls. The dotted line indicates the limit of detection. IgE, IgG2b, and IgG3 were undetectable in Atm −/− mice. Results are representative of two independent experiments. (E) Serum levels of Ag-specific IgA were measured in mice (five per group) immunized with TNP-KLH p.o., and serum was collected 4 d after the second immunization.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    32) Product Images from "Blood B Lymphocyte Stimulator (BLyS)/BAFF levels may reflect natural immunity to HIV in highly exposed uninfected Beninese Commercial Sex Workers"

    Article Title: Blood B Lymphocyte Stimulator (BLyS)/BAFF levels may reflect natural immunity to HIV in highly exposed uninfected Beninese Commercial Sex Workers

    Journal: Scientific Reports

    doi: 10.1038/srep32318

    Plasmablasts frequencies in the blood of HIV-uninfected non-commercial sex workers (CSWs), HIV-uninfected CSWs and HIV-infected CSWs. Analysis by flow-cytometry was performed as follows: ( A ) Cells were gated on live PBMCs and then on total B-cells (CD19 + CD3 − CD56 − ). Plasmablasts were gated on CD20 - CD38 ++ B-cells and further characterized as IgM + , IgA + or IgG + . Quadrants were set based on the expression values obtained with fluorescence minus one (FMO) and isotype controls. ( B ) The frequencies (%) of IgM + (left panel), IgA + (middle panel) and IgG + (right panel) plasmablasts are relative to total plasmablasts. The mean events gated for these populations are in the range of: CD19 + CD20 − CD38 ++ total plasmablasts (238 ± 112), IgM+ (25 ± 10), IgA+ (120 ± 50), IgG+ (45 ± 30). Representative FMO staining controls can be viewed in Supplementary Fig. S5 . Plasmablast percentages were compared with the Mann-Whitney U tests for pair-wise comparisons between HIV-uninfected CSWs and the two other groups. Significance levels are shown as *(p
    Figure Legend Snippet: Plasmablasts frequencies in the blood of HIV-uninfected non-commercial sex workers (CSWs), HIV-uninfected CSWs and HIV-infected CSWs. Analysis by flow-cytometry was performed as follows: ( A ) Cells were gated on live PBMCs and then on total B-cells (CD19 + CD3 − CD56 − ). Plasmablasts were gated on CD20 - CD38 ++ B-cells and further characterized as IgM + , IgA + or IgG + . Quadrants were set based on the expression values obtained with fluorescence minus one (FMO) and isotype controls. ( B ) The frequencies (%) of IgM + (left panel), IgA + (middle panel) and IgG + (right panel) plasmablasts are relative to total plasmablasts. The mean events gated for these populations are in the range of: CD19 + CD20 − CD38 ++ total plasmablasts (238 ± 112), IgM+ (25 ± 10), IgA+ (120 ± 50), IgG+ (45 ± 30). Representative FMO staining controls can be viewed in Supplementary Fig. S5 . Plasmablast percentages were compared with the Mann-Whitney U tests for pair-wise comparisons between HIV-uninfected CSWs and the two other groups. Significance levels are shown as *(p

    Techniques Used: Infection, Flow Cytometry, Cytometry, Expressing, Fluorescence, Staining, MANN-WHITNEY

    33) Product Images from "Towards an understanding of C9orf82 protein/CAAP1 function"

    Article Title: Towards an understanding of C9orf82 protein/CAAP1 function

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0210526

    Class switch recombination and proliferative potential of B cells lacking C9orf82 protein. (A) The class switch recombination potential as determined in vitro by switching of antigen inexperienced B cells to IgG3 and IgG1 after 4 days of exposure to LPS alone or LPS+IL-4, respectively is compared between wild type and C9orf82 ko/ko . (B) Proliferative potential of antigen inexperienced B cells after 3 days of exposure to LPS alone or LPS+IL-4 was measured by CFSE dilution and compared between the genotypes (a representation of 3 mice per genotype is shown).
    Figure Legend Snippet: Class switch recombination and proliferative potential of B cells lacking C9orf82 protein. (A) The class switch recombination potential as determined in vitro by switching of antigen inexperienced B cells to IgG3 and IgG1 after 4 days of exposure to LPS alone or LPS+IL-4, respectively is compared between wild type and C9orf82 ko/ko . (B) Proliferative potential of antigen inexperienced B cells after 3 days of exposure to LPS alone or LPS+IL-4 was measured by CFSE dilution and compared between the genotypes (a representation of 3 mice per genotype is shown).

    Techniques Used: In Vitro, Mouse Assay

    34) Product Images from "Low anti-RhD IgG-Fc-fucosylation in pregnancy: a new variable predicting severity in haemolytic disease of the fetus and newborn"

    Article Title: Low anti-RhD IgG-Fc-fucosylation in pregnancy: a new variable predicting severity in haemolytic disease of the fetus and newborn

    Journal: British Journal of Haematology

    doi: 10.1111/bjh.12965

    IgG1 glycosylation profiles of a pregnant woman with anti-D. Tryptic IgG1 Fc glycopeptides of total IgG1 (A) and anti-D specific IgG1 (B), affinity-purified from the serum of a pregnant woman, were measured by positive-ion reflectron mode matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The profile shown represents specific antibodies with a low fucosylation of 16%, while the total IgG1 was highly fucosylated (94%). For the assignment and the definitions of the glycopeptides signals see Fig 1 . Pep, peptide moiety; *, contaminant.
    Figure Legend Snippet: IgG1 glycosylation profiles of a pregnant woman with anti-D. Tryptic IgG1 Fc glycopeptides of total IgG1 (A) and anti-D specific IgG1 (B), affinity-purified from the serum of a pregnant woman, were measured by positive-ion reflectron mode matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The profile shown represents specific antibodies with a low fucosylation of 16%, while the total IgG1 was highly fucosylated (94%). For the assignment and the definitions of the glycopeptides signals see Fig 1 . Pep, peptide moiety; *, contaminant.

    Techniques Used: Affinity Purification, Mass Spectrometry

    Low anti-D IgG1-fucosylation induces stronger NK-cell mediated ADCC. Natural killer (NK)-cell ADCC versus the levels of galactosylation (A), bisection (B) and fucosylation (C) found in IgG1 anti-D. For the NK-cell ADCC only samples containing enough material and with an anti-D titre of 1/128 were plotted. Statistical analyses were performed using two-tailed (A, B) or one-tailed (C) Pearson correlation with significance set at P = 0·05.
    Figure Legend Snippet: Low anti-D IgG1-fucosylation induces stronger NK-cell mediated ADCC. Natural killer (NK)-cell ADCC versus the levels of galactosylation (A), bisection (B) and fucosylation (C) found in IgG1 anti-D. For the NK-cell ADCC only samples containing enough material and with an anti-D titre of 1/128 were plotted. Statistical analyses were performed using two-tailed (A, B) or one-tailed (C) Pearson correlation with significance set at P = 0·05.

    Techniques Used: Two Tailed Test, One-tailed Test

    Anti-D IgG1 in pregnancy displays pronounced lowered Fc-fucosylation. Relative expression levels of major IgG-Fc Asn-297 glycoforms for both total IgG1 ( x -axis) and antigen-specific IgG1 ( y -axis) for 70 pregnancy-induced anti-D serum samples (A–C). Serum populations were analysed for Fc-galactosylation (A), Fc-bisection (bisecting N -acetylglucosamine) (B) and Fc-fucosylation (C). The statistical outcome between two-tailed paired t -test analysis of total IgG1 versus . specific antibodies is listed in each panel. The diagonal, dotted lines represent the theoretical equal glycosylation of total IgG1 and anti-D IgG1. Normal total IgG1 values (non-pregnancy setting): galactose (61·92%), bisection (13·80%) and fucose (92·86%) ( Selman et al , 2012 ).
    Figure Legend Snippet: Anti-D IgG1 in pregnancy displays pronounced lowered Fc-fucosylation. Relative expression levels of major IgG-Fc Asn-297 glycoforms for both total IgG1 ( x -axis) and antigen-specific IgG1 ( y -axis) for 70 pregnancy-induced anti-D serum samples (A–C). Serum populations were analysed for Fc-galactosylation (A), Fc-bisection (bisecting N -acetylglucosamine) (B) and Fc-fucosylation (C). The statistical outcome between two-tailed paired t -test analysis of total IgG1 versus . specific antibodies is listed in each panel. The diagonal, dotted lines represent the theoretical equal glycosylation of total IgG1 and anti-D IgG1. Normal total IgG1 values (non-pregnancy setting): galactose (61·92%), bisection (13·80%) and fucose (92·86%) ( Selman et al , 2012 ).

    Techniques Used: Expressing, Two Tailed Test

    Anti-D IgG1 with lower core fucosylation induces more severe haemolysis. No significant correlation was found between anti-D titre and Haemoglobin levels (A). However, the degree of IgG1-anti-D fucosylation in pregnancy correlated significantly with fetal or neonatal haemoglobin levels (B). Statistical analyses were performed using one-tailed Pearson correlation with significance set at P = 0·05. NS: not significant.
    Figure Legend Snippet: Anti-D IgG1 with lower core fucosylation induces more severe haemolysis. No significant correlation was found between anti-D titre and Haemoglobin levels (A). However, the degree of IgG1-anti-D fucosylation in pregnancy correlated significantly with fetal or neonatal haemoglobin levels (B). Statistical analyses were performed using one-tailed Pearson correlation with significance set at P = 0·05. NS: not significant.

    Techniques Used: One-tailed Test

    35) Product Images from "Naloxone/alum mixture a potent adjuvant for HIV-1 vaccine: induction of cellular and poly-isotypic humoral immune responses"

    Article Title: Naloxone/alum mixture a potent adjuvant for HIV-1 vaccine: induction of cellular and poly-isotypic humoral immune responses

    Journal: Pathogens and Global Health

    doi: 10.1179/2047773215Y.0000000035

    Detection of IgG1 (a), IgG2a (b), IgG2b (c), IgG3 (d) and IgM, (e) specific antibodies. The measurements were made with sera of experimental mice, and the values represent the means of optical density ± standard deviation. Asterisks indicate the groups which were significantly different and ND indicates not detectable differences ( p
    Figure Legend Snippet: Detection of IgG1 (a), IgG2a (b), IgG2b (c), IgG3 (d) and IgM, (e) specific antibodies. The measurements were made with sera of experimental mice, and the values represent the means of optical density ± standard deviation. Asterisks indicate the groups which were significantly different and ND indicates not detectable differences ( p

    Techniques Used: Mouse Assay, Standard Deviation

    36) Product Images from "Neurofascin-155 IgG4 in chronic inflammatory demyelinating polyneuropathy"

    Article Title: Neurofascin-155 IgG4 in chronic inflammatory demyelinating polyneuropathy

    Journal: Neurology

    doi: 10.1212/WNL.0000000000002418

    Anti-NF155 IgG antibodies target an N -glycosylated module
    Figure Legend Snippet: Anti-NF155 IgG antibodies target an N -glycosylated module

    Techniques Used:

    Clinical features of anti-NF155 IgG4-positive CIDP.
    Figure Legend Snippet: Clinical features of anti-NF155 IgG4-positive CIDP.

    Techniques Used:

    37) Product Images from "Utilization of Immunoglobulin G Fc Receptors by Human Immunodeficiency Virus Type 1: a Specific Role for Antibodies against the Membrane-Proximal External Region of gp41 ▿"

    Article Title: Utilization of Immunoglobulin G Fc Receptors by Human Immunodeficiency Virus Type 1: a Specific Role for Antibodies against the Membrane-Proximal External Region of gp41 ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00656-09

    Importance of IgG subclass and the Fc region of Ab. Left panel, neutralization curves for 2F5 (IgG1) and 4E10 (IgG1) assayed against two HIV-1 subtype B Env-pseudotyped viruses in parental TZM-bl cells (squares) and in TZM-bl cells expressing either FcγRI (circles) or FcγRIIb (triangles). Right panel, neutralization potencies of 2F5 IgG1 and 4E10 IgG1 (black bars), 2F5 IgG3 and 4E10 IgG3 (light gray bars), and 4E10 Fab (dark gray bars) in parental TZM-bl cells and TZM-bl cells expressing either FcγRI, FcγRIIa, FcγRIIb, or FcγRIIIa. ID50, 50% inhibitory dose.
    Figure Legend Snippet: Importance of IgG subclass and the Fc region of Ab. Left panel, neutralization curves for 2F5 (IgG1) and 4E10 (IgG1) assayed against two HIV-1 subtype B Env-pseudotyped viruses in parental TZM-bl cells (squares) and in TZM-bl cells expressing either FcγRI (circles) or FcγRIIb (triangles). Right panel, neutralization potencies of 2F5 IgG1 and 4E10 IgG1 (black bars), 2F5 IgG3 and 4E10 IgG3 (light gray bars), and 4E10 Fab (dark gray bars) in parental TZM-bl cells and TZM-bl cells expressing either FcγRI, FcγRIIa, FcγRIIb, or FcγRIIIa. ID50, 50% inhibitory dose.

    Techniques Used: Neutralization, Expressing

    Relative binding of HIV-1 broadly neutralizing monoclonal Abs to FcγRI on TZM-bl cells. TZM-bl cells expressing FcγRI were incubated with either IgG1b12, 2G12, 2F5, or 4E10 for 1 h at 4°C. Cells were washed three times, stained with FITC-conjugated goat anti-human IgG (Fab specific), and analyzed by flow cytometry. Top, percentage of cells staining positive; bottom, mean fluorescence intensity (MFI) of positive cells. Background staining with an isotype control Ab was subtracted in both cases.
    Figure Legend Snippet: Relative binding of HIV-1 broadly neutralizing monoclonal Abs to FcγRI on TZM-bl cells. TZM-bl cells expressing FcγRI were incubated with either IgG1b12, 2G12, 2F5, or 4E10 for 1 h at 4°C. Cells were washed three times, stained with FITC-conjugated goat anti-human IgG (Fab specific), and analyzed by flow cytometry. Top, percentage of cells staining positive; bottom, mean fluorescence intensity (MFI) of positive cells. Background staining with an isotype control Ab was subtracted in both cases.

    Techniques Used: Binding Assay, Expressing, Incubation, Staining, Flow Cytometry, Cytometry, Fluorescence

    Normal human serum abrogates the FcγRI effect on 2F5 and 4E10. Neutralization assays were performed with HIV-1 700010040.C9.4520 in parental TZM-bl cells and in TZM-bl cells expressing either FcγRI or FcγRIIb in the absence of serum (black bars) or in the presence of either 5% fresh normal human serum (light gray bars) or 5% heat-inactivated normal human serum (dark gray bars). The monoclonal Abs were of the IgG1 subclass. ID50, 50% inhibitory dose.
    Figure Legend Snippet: Normal human serum abrogates the FcγRI effect on 2F5 and 4E10. Neutralization assays were performed with HIV-1 700010040.C9.4520 in parental TZM-bl cells and in TZM-bl cells expressing either FcγRI or FcγRIIb in the absence of serum (black bars) or in the presence of either 5% fresh normal human serum (light gray bars) or 5% heat-inactivated normal human serum (dark gray bars). The monoclonal Abs were of the IgG1 subclass. ID50, 50% inhibitory dose.

    Techniques Used: Neutralization, Expressing

    Effect of FcγR expression on HIV-1 neutralization by broadly neutralizing monoclonal Abs and HIVIG. The neutralizing activities of monoclonal Abs b12, 2G12, 2F5, and 4E10 (all IgG1) and HIVIG (purified IgG fraction prepared from pooled HIV-1-positive plasma samples) were assessed against four strains each of HIV-1 subtype B (6535.3, QH0692.42, SC05.8C11.2344, and SC422661.87), subtype C (Du156.12, Du422.1, ZM197M.PB7, and CAP45.2.00.G3), and subtype A (Q23.17, Q259.d2.17, Q461.a2, and Q769.d22) in parental TZM-bl cells (black bars) and in TZM-bl cells expressing FcγR (gray bars). ID50, 50% inhibitory dose.
    Figure Legend Snippet: Effect of FcγR expression on HIV-1 neutralization by broadly neutralizing monoclonal Abs and HIVIG. The neutralizing activities of monoclonal Abs b12, 2G12, 2F5, and 4E10 (all IgG1) and HIVIG (purified IgG fraction prepared from pooled HIV-1-positive plasma samples) were assessed against four strains each of HIV-1 subtype B (6535.3, QH0692.42, SC05.8C11.2344, and SC422661.87), subtype C (Du156.12, Du422.1, ZM197M.PB7, and CAP45.2.00.G3), and subtype A (Q23.17, Q259.d2.17, Q461.a2, and Q769.d22) in parental TZM-bl cells (black bars) and in TZM-bl cells expressing FcγR (gray bars). ID50, 50% inhibitory dose.

    Techniques Used: Expressing, Neutralization, Purification

    38) Product Images from "Interleukin-6 Receptor Blockade Selectively Reduces IL-21 Production by CD4 T Cells and IgG4 Autoantibodies in Rheumatoid Arthritis"

    Article Title: Interleukin-6 Receptor Blockade Selectively Reduces IL-21 Production by CD4 T Cells and IgG4 Autoantibodies in Rheumatoid Arthritis

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.5996

    Effect of tocilizumab on IgG isotypes and IgG4 autoantibodies. (A) and (B) Serum levels of total IgG2 ( A ) and IgG4 ( B ) prior to (0 months) and 6 months after the treatment with tocilizumab in the eight patients. ( C ) IgG4-specific anti-CCP Ab levels in serum. Six patients (Patients #1, 2, 4, 5, 6 and 7) had detectable levels of IgG4-anti-CCP Abs in serum prior to the initiation of tocilizumab treatment (0 months). ( D ) IgG1-specific anti-CCP Ab levels in serum. Five patients (Patients #1, 2, 4, 6 and 7) showed detectable levels of IgG1-anti-CCP Abs prior to the treatment. ( E ) Fold reduction in the serum levels of IgG1-specific anti-CCP Abs and IgG4-specific anti-CCP Abs between 0 months (prior to the treatment) and 6 months after the initiation of the treatment. Results of the repeated measures analysis of variance suggest that the fold reduction in IgG1 differs from that observed in IgG4 (p=0.011 for the interaction effect). Follow-up tests of simple effects shows a statistically significant (p =0.011) fold-reduction in IgG4 anti-CCP Abs (denoted by *) while there was no reduction in IgG1 anti-CCP Abs levels (p=0.185). Note that a fold-reduction equal to 1 between 0 months and 6 months means no effect on IgG levels with treatment. ( F ) Purified B cells from healthy volunteers (n=4) were activated in vitro with CD40L-expressing cells in the presence of medium, IL-4 or IL-21. The levels of IgG4 in the supernatants were determined after 6 days. Fold induction for each subject in the levels of IgG4 produced by B cells activated with IL-4 or IL-21 relative to the levels by B cells activated with just medium is shown. The statistically significant difference between fold-induction obtained with IL-21 relative to IL-4 Abs was determined by paired t test analysis, p =0.0206.
    Figure Legend Snippet: Effect of tocilizumab on IgG isotypes and IgG4 autoantibodies. (A) and (B) Serum levels of total IgG2 ( A ) and IgG4 ( B ) prior to (0 months) and 6 months after the treatment with tocilizumab in the eight patients. ( C ) IgG4-specific anti-CCP Ab levels in serum. Six patients (Patients #1, 2, 4, 5, 6 and 7) had detectable levels of IgG4-anti-CCP Abs in serum prior to the initiation of tocilizumab treatment (0 months). ( D ) IgG1-specific anti-CCP Ab levels in serum. Five patients (Patients #1, 2, 4, 6 and 7) showed detectable levels of IgG1-anti-CCP Abs prior to the treatment. ( E ) Fold reduction in the serum levels of IgG1-specific anti-CCP Abs and IgG4-specific anti-CCP Abs between 0 months (prior to the treatment) and 6 months after the initiation of the treatment. Results of the repeated measures analysis of variance suggest that the fold reduction in IgG1 differs from that observed in IgG4 (p=0.011 for the interaction effect). Follow-up tests of simple effects shows a statistically significant (p =0.011) fold-reduction in IgG4 anti-CCP Abs (denoted by *) while there was no reduction in IgG1 anti-CCP Abs levels (p=0.185). Note that a fold-reduction equal to 1 between 0 months and 6 months means no effect on IgG levels with treatment. ( F ) Purified B cells from healthy volunteers (n=4) were activated in vitro with CD40L-expressing cells in the presence of medium, IL-4 or IL-21. The levels of IgG4 in the supernatants were determined after 6 days. Fold induction for each subject in the levels of IgG4 produced by B cells activated with IL-4 or IL-21 relative to the levels by B cells activated with just medium is shown. The statistically significant difference between fold-induction obtained with IL-21 relative to IL-4 Abs was determined by paired t test analysis, p =0.0206.

    Techniques Used: Purification, In Vitro, Expressing, Produced

    39) Product Images from "Unusual Patterns of IgG Avidity in Some Young Children following Two Doses of the Adjuvanted Pandemic H1N1 (2009) Influenza Virus Vaccine"

    Article Title: Unusual Patterns of IgG Avidity in Some Young Children following Two Doses of the Adjuvanted Pandemic H1N1 (2009) Influenza Virus Vaccine

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00619-12

    Influenza virus-specific IgG subclass analysis in young children immunized with two doses of AS03-adjuvanted pdmH1N1 split vaccine. Serum samples were collected before and 3 weeks after each immunization, which were given at visit 1 and visit 2. The concentrations
    Figure Legend Snippet: Influenza virus-specific IgG subclass analysis in young children immunized with two doses of AS03-adjuvanted pdmH1N1 split vaccine. Serum samples were collected before and 3 weeks after each immunization, which were given at visit 1 and visit 2. The concentrations

    Techniques Used:

    40) Product Images from "Malaria vaccine candidate based on Duffy-binding protein elicits strain transcending functional antibodies in a Phase I trial"

    Article Title: Malaria vaccine candidate based on Duffy-binding protein elicits strain transcending functional antibodies in a Phase I trial

    Journal: NPJ Vaccines

    doi: 10.1038/s41541-018-0083-3

    Humoral response to immunization with PvDBPII formulated with GLA-SE. a Kinetics of PvDBPII-specific Geometric Mean antibody titres for all Study Groups (PP Population). Geometric mean antibody levels (U/mL) to PvDBPII measured by ELISA in sera collected from groups immunized with 10, 25, or 50 μg PvDBPII/GLA-SE and Hepatitis B vaccine. Study participants were immunized on Days 0, 28 and 56, and sera were collected on Days 0, 28, 56, 84 and 180. Antibody levels measured by ELISA were expressed as Geometric Mean Titres (GMT) with 95% confidence interval for the nine subjects. b Serum IgG subclass responses to PvDBPII in three PvDBPII dose groups (PP Population). Geometric means with SD are shown
    Figure Legend Snippet: Humoral response to immunization with PvDBPII formulated with GLA-SE. a Kinetics of PvDBPII-specific Geometric Mean antibody titres for all Study Groups (PP Population). Geometric mean antibody levels (U/mL) to PvDBPII measured by ELISA in sera collected from groups immunized with 10, 25, or 50 μg PvDBPII/GLA-SE and Hepatitis B vaccine. Study participants were immunized on Days 0, 28 and 56, and sera were collected on Days 0, 28, 56, 84 and 180. Antibody levels measured by ELISA were expressed as Geometric Mean Titres (GMT) with 95% confidence interval for the nine subjects. b Serum IgG subclass responses to PvDBPII in three PvDBPII dose groups (PP Population). Geometric means with SD are shown

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Related Articles

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    Article Snippet: .. For IgG subclass ELISA analysis, goat anti-mouse IgG1, IgG2a, IgG2b, and IgG3 (heavy chain specific; Sigma), and alkaline phosphatase-conjugated rabbit anti-goat IgG (whole molecule; Sigma) were used. ..

    Purification:

    Article Title: Fabrication of a Dual Substrate Display to Test Roles of Cell Adhesion Proteins in Vesicle Targeting to Plasma Membrane Domains
    Article Snippet: .. Where appropriate, purified IgG heavy chain (Fc; 200μg/ml; Sigma) was substituted in the final step in place of Fc-E-cadherin protein as a negative (non-adhesive) control. .. A Madin Darby canine kidney (MDCK) IIG cell line stably expressing VSVG-YFP, a marker for basolateral exocytosis, was generated using Lipofectamine 2000 (Invitrogen) transfection, Fluorescent Activated Cell Sorting flow cytometry, followed by standard cell cloning selection procedures.

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    Millipore goat anti human igg
    RTS,S vaccine-induced immunity declines over time. A random selection of children vaccinated with RTS,S (Manhiça cohort, n = 30) was tested for C1q-fixation ( a ), <t>IgG/IgM</t> ( b ), and IgG subclasses ( c ) to CSP at months 3, 8.5, 21, 33, 45, and 6. Note that due to low reactivity, C1q-fixation was re-tested at a higher dilution of 1/110, in addition to 1/250, to confirm results. Samples were tested in duplicate, and the median and 95% CI of the median from each time point group are shown by the symbol and shaded area, respectively
    Goat Anti Human Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore cy3 conjugated anti rabbit igg
    Localization of FGFRs in human sperm. Sperm cells were stained with anti FGFR1, FGFR2, FGFR3 and FGFR4 or rabbit <t>IgG</t> and a secondary antibody labeled with <t>Cy3.</t> The corresponding fields stained with FITC-PSA to assess acrosomal status are shown. Bar: 10 μm. On the right, a representative image of individual sperm is depicted; ( A ) sperm stained with anti FGFR antibody and Cy3-conjugated secondary antibody, ( B ) FITC-PSA, ( C ) merge.
    Cy3 Conjugated Anti Rabbit Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore control igg antibody
    Robo1 signaling activates Rho GTPases in transformed cells. Src transformed cells were treated with <t>R5</t> antibody or control antiserum <t>(IgG)</t> and examined for total and activated Cdc42 and Rac1 GTPases in panels a and b , respectively. Western blotting was performed to detect active (GTP bound) Cdc42 and Rac1, total Cdc42 and Rac1, and GST. Levels of active Cdc42 and Rac1 were quantitated and shown as the percent of untreated control cells (mean+SEM, n=3). Experiments were performed with Cx43Ko and wild type cells, with results from Cx43Ko cells shown. Single and triple asterisks indicate p values less than 0.5 and 0.005, respectively (by t-test).
    Control Igg Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore igg2
    <t>IgG-,</t> Fab-, and Fcγ-binding activities of recombinant TspB derivatives and live bacteria. (A) Schematic representation of the putative domains of TspB and corresponding recombinant proteins. (B) TspB NT (lane 1) and CT (lane 2) derivatives resolved
    Igg2, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    RTS,S vaccine-induced immunity declines over time. A random selection of children vaccinated with RTS,S (Manhiça cohort, n = 30) was tested for C1q-fixation ( a ), IgG/IgM ( b ), and IgG subclasses ( c ) to CSP at months 3, 8.5, 21, 33, 45, and 6. Note that due to low reactivity, C1q-fixation was re-tested at a higher dilution of 1/110, in addition to 1/250, to confirm results. Samples were tested in duplicate, and the median and 95% CI of the median from each time point group are shown by the symbol and shaded area, respectively

    Journal: BMC Medicine

    Article Title: Induction and decay of functional complement-fixing antibodies by the RTS,S malaria vaccine in children, and a negative impact of malaria exposure

    doi: 10.1186/s12916-019-1277-x

    Figure Lengend Snippet: RTS,S vaccine-induced immunity declines over time. A random selection of children vaccinated with RTS,S (Manhiça cohort, n = 30) was tested for C1q-fixation ( a ), IgG/IgM ( b ), and IgG subclasses ( c ) to CSP at months 3, 8.5, 21, 33, 45, and 6. Note that due to low reactivity, C1q-fixation was re-tested at a higher dilution of 1/110, in addition to 1/250, to confirm results. Samples were tested in duplicate, and the median and 95% CI of the median from each time point group are shown by the symbol and shaded area, respectively

    Article Snippet: Plates were washed, and antibody isotypes were detected using goat anti-human IgG and IgM conjugated to horseradish peroxidase (HRP, Millipore, Burlington, USA) at 1/2500 dilution in buffer for 1 h at RT.

    Techniques: Selection

    RTS,S vaccine-induced IgG and IgM antibodies to CSP. Children in RTS,S and comparator vaccine groups from Manhiça (black box plots; N = 50 and N = 25, respectively) and Ilha Josina cohorts (gray box plots; N = 49 and N = 24, respectively) were tested for IgG ( a ), IgG subclasses ( b ), and IgM ( c ) to CSP. Sera collected at baseline (month 0, M0) and after vaccination (month 3, M3) were tested in duplicate (note that only M3 was tested in b ), and the mean value was used to generate box plots whereby top, center, and bottom horizontal lines represent the 75th percentile, median, and 25th percentile, respectively; upper and lower whiskers represent the highest and lowest values within 1.5× IQR, respectively; and values that exceed this range are presented as dots. Malaria-naïve negative controls from Melbourne donors were used to calculate the seropositivity cutoff value (dashed lines), and the percentage of individuals above this threshold are shown. Reactivity between paired samples and unpaired samples were compared using Wilcoxon matched-pairs signed-rank test and Mann-Whitney U test, respectively

    Journal: BMC Medicine

    Article Title: Induction and decay of functional complement-fixing antibodies by the RTS,S malaria vaccine in children, and a negative impact of malaria exposure

    doi: 10.1186/s12916-019-1277-x

    Figure Lengend Snippet: RTS,S vaccine-induced IgG and IgM antibodies to CSP. Children in RTS,S and comparator vaccine groups from Manhiça (black box plots; N = 50 and N = 25, respectively) and Ilha Josina cohorts (gray box plots; N = 49 and N = 24, respectively) were tested for IgG ( a ), IgG subclasses ( b ), and IgM ( c ) to CSP. Sera collected at baseline (month 0, M0) and after vaccination (month 3, M3) were tested in duplicate (note that only M3 was tested in b ), and the mean value was used to generate box plots whereby top, center, and bottom horizontal lines represent the 75th percentile, median, and 25th percentile, respectively; upper and lower whiskers represent the highest and lowest values within 1.5× IQR, respectively; and values that exceed this range are presented as dots. Malaria-naïve negative controls from Melbourne donors were used to calculate the seropositivity cutoff value (dashed lines), and the percentage of individuals above this threshold are shown. Reactivity between paired samples and unpaired samples were compared using Wilcoxon matched-pairs signed-rank test and Mann-Whitney U test, respectively

    Article Snippet: Plates were washed, and antibody isotypes were detected using goat anti-human IgG and IgM conjugated to horseradish peroxidase (HRP, Millipore, Burlington, USA) at 1/2500 dilution in buffer for 1 h at RT.

    Techniques: MANN-WHITNEY

    High variability among RTS,S vaccine-induced IgG targeting the central repeat and C-terminal regions of CSP. Children in RTS,S vaccine group from Manhiça and Ilha Josina cohorts ( N = 99, 3 M) were tested for IgG to NANP and CT and C1q-fixation to CSP, and the values were used for the following analysis. a Heat map of children arranged in descending (top to bottom) order of C1q-fixation (left), corresponding IgG to NANP and CT (middle), and for comparison IgM to NANP and CT (right). b Variability between epitope specificity was quantified by calculating the ratio of NANP-to-CT IgG. Children with low variability were considered to have equal reactivity to NANP and CT (ratio between 0.75 and 1.25 shown in white), and children exceeding this range were considered to have a NANP- or CT-skewed response (ratio > 1.25 shown in purple and ratio

    Journal: BMC Medicine

    Article Title: Induction and decay of functional complement-fixing antibodies by the RTS,S malaria vaccine in children, and a negative impact of malaria exposure

    doi: 10.1186/s12916-019-1277-x

    Figure Lengend Snippet: High variability among RTS,S vaccine-induced IgG targeting the central repeat and C-terminal regions of CSP. Children in RTS,S vaccine group from Manhiça and Ilha Josina cohorts ( N = 99, 3 M) were tested for IgG to NANP and CT and C1q-fixation to CSP, and the values were used for the following analysis. a Heat map of children arranged in descending (top to bottom) order of C1q-fixation (left), corresponding IgG to NANP and CT (middle), and for comparison IgM to NANP and CT (right). b Variability between epitope specificity was quantified by calculating the ratio of NANP-to-CT IgG. Children with low variability were considered to have equal reactivity to NANP and CT (ratio between 0.75 and 1.25 shown in white), and children exceeding this range were considered to have a NANP- or CT-skewed response (ratio > 1.25 shown in purple and ratio

    Article Snippet: Plates were washed, and antibody isotypes were detected using goat anti-human IgG and IgM conjugated to horseradish peroxidase (HRP, Millipore, Burlington, USA) at 1/2500 dilution in buffer for 1 h at RT.

    Techniques:

    Functional complement-fixing antibodies target the central repeat and C-terminal regions of CSP. Children in RTS,S vaccine group from Manhiça (black box plots; N = 50) and Ilha Josina cohorts (gray box plots; N = 49) were tested for IgG ( a ) and IgM ( b ) to NANP and CT regions of CSP. Sera collected at baseline (month 0, M0) and after vaccination (month 3, M3) were tested in duplicate, and the mean value was used to generate box plots whereby top, center, and bottom horizontal lines represent the 75th percentile, median, and 25th percentile, respectively; upper and lower whiskers represent the highest and lowest values within 1.5× IQR, respectively; and values that exceed this range are presented as dots. Malaria-naïve negative controls from Melbourne donors were used to calculate the seropositivity cutoff values (dashed lines), and the percentages of individuals above this threshold are shown. Reactivity between paired samples was compared using Wilcoxon matched-pairs signed-rank test. c , d Children in RTS,S vaccine group from Manhiça and Ilha Josina cohorts ( N = 99, M3) were tested for C1q-fixation to CSP, NANP, and CT, and the values were plotted compared to IgG reactivity (c) and C1q fixation to CSP, NANP, and CT were correlated (d)

    Journal: BMC Medicine

    Article Title: Induction and decay of functional complement-fixing antibodies by the RTS,S malaria vaccine in children, and a negative impact of malaria exposure

    doi: 10.1186/s12916-019-1277-x

    Figure Lengend Snippet: Functional complement-fixing antibodies target the central repeat and C-terminal regions of CSP. Children in RTS,S vaccine group from Manhiça (black box plots; N = 50) and Ilha Josina cohorts (gray box plots; N = 49) were tested for IgG ( a ) and IgM ( b ) to NANP and CT regions of CSP. Sera collected at baseline (month 0, M0) and after vaccination (month 3, M3) were tested in duplicate, and the mean value was used to generate box plots whereby top, center, and bottom horizontal lines represent the 75th percentile, median, and 25th percentile, respectively; upper and lower whiskers represent the highest and lowest values within 1.5× IQR, respectively; and values that exceed this range are presented as dots. Malaria-naïve negative controls from Melbourne donors were used to calculate the seropositivity cutoff values (dashed lines), and the percentages of individuals above this threshold are shown. Reactivity between paired samples was compared using Wilcoxon matched-pairs signed-rank test. c , d Children in RTS,S vaccine group from Manhiça and Ilha Josina cohorts ( N = 99, M3) were tested for C1q-fixation to CSP, NANP, and CT, and the values were plotted compared to IgG reactivity (c) and C1q fixation to CSP, NANP, and CT were correlated (d)

    Article Snippet: Plates were washed, and antibody isotypes were detected using goat anti-human IgG and IgM conjugated to horseradish peroxidase (HRP, Millipore, Burlington, USA) at 1/2500 dilution in buffer for 1 h at RT.

    Techniques: Functional Assay

    Localization of FGFRs in human sperm. Sperm cells were stained with anti FGFR1, FGFR2, FGFR3 and FGFR4 or rabbit IgG and a secondary antibody labeled with Cy3. The corresponding fields stained with FITC-PSA to assess acrosomal status are shown. Bar: 10 μm. On the right, a representative image of individual sperm is depicted; ( A ) sperm stained with anti FGFR antibody and Cy3-conjugated secondary antibody, ( B ) FITC-PSA, ( C ) merge.

    Journal: PLoS ONE

    Article Title: Fibroblast Growth Factor Receptors (FGFRs) in Human Sperm: Expression, Functionality and Involvement in Motility Regulation

    doi: 10.1371/journal.pone.0127297

    Figure Lengend Snippet: Localization of FGFRs in human sperm. Sperm cells were stained with anti FGFR1, FGFR2, FGFR3 and FGFR4 or rabbit IgG and a secondary antibody labeled with Cy3. The corresponding fields stained with FITC-PSA to assess acrosomal status are shown. Bar: 10 μm. On the right, a representative image of individual sperm is depicted; ( A ) sperm stained with anti FGFR antibody and Cy3-conjugated secondary antibody, ( B ) FITC-PSA, ( C ) merge.

    Article Snippet: Other antibodies used were: anti pFGFR Tyr653/654 (#3476, Cell Signaling Technology, Inc., Beverley, MA, USA), anti pERK (sc-7383, Santa Cruz, and #4370, Cell Signaling), anti ERK (sc-94, Santa Cruz), anti pAkt Ser473 (sc-7985, Santa Cruz, and #4060, Cell Signaling), anti Akt (#4691, Cell Signaling), rabbit immunoglobulin G (IgG) (Sigma), horse-radish peroxidase (HRP)-conjugated anti-rabbit IgG (Sigma), Cy3-conjugated anti-rabbit IgG and FITC-conjugated anti-mouse IgG (Chemicon-Millipore, Billerica, MA, USA) and anti-rabbit IgG (Sigma).

    Techniques: Staining, Labeling

    Robo1 signaling activates Rho GTPases in transformed cells. Src transformed cells were treated with R5 antibody or control antiserum (IgG) and examined for total and activated Cdc42 and Rac1 GTPases in panels a and b , respectively. Western blotting was performed to detect active (GTP bound) Cdc42 and Rac1, total Cdc42 and Rac1, and GST. Levels of active Cdc42 and Rac1 were quantitated and shown as the percent of untreated control cells (mean+SEM, n=3). Experiments were performed with Cx43Ko and wild type cells, with results from Cx43Ko cells shown. Single and triple asterisks indicate p values less than 0.5 and 0.005, respectively (by t-test).

    Journal: Oncotarget

    Article Title: Src activates Abl to augment Robo1 expression in order to promote tumor cell migration

    doi:

    Figure Lengend Snippet: Robo1 signaling activates Rho GTPases in transformed cells. Src transformed cells were treated with R5 antibody or control antiserum (IgG) and examined for total and activated Cdc42 and Rac1 GTPases in panels a and b , respectively. Western blotting was performed to detect active (GTP bound) Cdc42 and Rac1, total Cdc42 and Rac1, and GST. Levels of active Cdc42 and Rac1 were quantitated and shown as the percent of untreated control cells (mean+SEM, n=3). Experiments were performed with Cx43Ko and wild type cells, with results from Cx43Ko cells shown. Single and triple asterisks indicate p values less than 0.5 and 0.005, respectively (by t-test).

    Article Snippet: For some experiments, cells were treated overnight with 50 μg/ml cyclohexamide (Sigma, C7698), 60 nM R5 antibody to the extracellular region of Robo1, 60 nM control IgG antibody [ , ], or 40 μM GNF-2 Abl kinase inhibitor (Calbiochem, 197221).

    Techniques: Transformation Assay, Western Blot

    Src utilizes Robo1 to promote cell migration. Cells obtained from wild type (WT) or homozygous null Cx43 knockout (Cx43Ko) mouse embryos were transfected with v-Src or the empty parental vector and plated (10,000 per well) were plated on standard or ultra low attachment culture dishes to evaluate (a) anchored and (b) nonanchored cell growth. Cell numbers were determined by Coulter counter at the indicated time points for anchored cells or at 7 days for nonanchored cells. Data are shown as number of cells per well at the indicated time points (mean+SEM, n=3). (c) Cell migration was examined by a wound healing assay on nontransformed cells and Src transformed cells treated with IgG control antiserum, R5 antiserum to block Robo1 activity, or GNF-2 Abl kinase blocker. Migration was quantitated as the number of cells that entered a 1.8 mm2 area of the wound during 24 hours (mean+SEM, n=5). (d) Src transformed cells were treated with R5 antibody or control antiserum and analyzed by Western blotting for Robo1, active N-WASP (p-N-WASP), or β-actin. N-WASP activity was then quantitated and shown as percent of untreated control cells (mean+SEM, n=2). Experiments were performed with Cx43Ko and WT cells, with results from Cx43Ko cells shown in panel d. Double and triple asterisk indicate p values less than 0.01 and 0.005 compared to controls, respectively (by t-test).

    Journal: Oncotarget

    Article Title: Src activates Abl to augment Robo1 expression in order to promote tumor cell migration

    doi:

    Figure Lengend Snippet: Src utilizes Robo1 to promote cell migration. Cells obtained from wild type (WT) or homozygous null Cx43 knockout (Cx43Ko) mouse embryos were transfected with v-Src or the empty parental vector and plated (10,000 per well) were plated on standard or ultra low attachment culture dishes to evaluate (a) anchored and (b) nonanchored cell growth. Cell numbers were determined by Coulter counter at the indicated time points for anchored cells or at 7 days for nonanchored cells. Data are shown as number of cells per well at the indicated time points (mean+SEM, n=3). (c) Cell migration was examined by a wound healing assay on nontransformed cells and Src transformed cells treated with IgG control antiserum, R5 antiserum to block Robo1 activity, or GNF-2 Abl kinase blocker. Migration was quantitated as the number of cells that entered a 1.8 mm2 area of the wound during 24 hours (mean+SEM, n=5). (d) Src transformed cells were treated with R5 antibody or control antiserum and analyzed by Western blotting for Robo1, active N-WASP (p-N-WASP), or β-actin. N-WASP activity was then quantitated and shown as percent of untreated control cells (mean+SEM, n=2). Experiments were performed with Cx43Ko and WT cells, with results from Cx43Ko cells shown in panel d. Double and triple asterisk indicate p values less than 0.01 and 0.005 compared to controls, respectively (by t-test).

    Article Snippet: For some experiments, cells were treated overnight with 50 μg/ml cyclohexamide (Sigma, C7698), 60 nM R5 antibody to the extracellular region of Robo1, 60 nM control IgG antibody [ , ], or 40 μM GNF-2 Abl kinase inhibitor (Calbiochem, 197221).

    Techniques: Migration, Knock-Out, Transfection, Plasmid Preparation, Wound Healing Assay, Transformation Assay, Blocking Assay, Activity Assay, Western Blot

    IgG-, Fab-, and Fcγ-binding activities of recombinant TspB derivatives and live bacteria. (A) Schematic representation of the putative domains of TspB and corresponding recombinant proteins. (B) TspB NT (lane 1) and CT (lane 2) derivatives resolved

    Journal: Infection and Immunity

    Article Title: Resistance of Neisseria meningitidis to Human Serum Depends on T and B Cell Stimulating Protein B

    doi: 10.1128/IAI.03134-14

    Figure Lengend Snippet: IgG-, Fab-, and Fcγ-binding activities of recombinant TspB derivatives and live bacteria. (A) Schematic representation of the putative domains of TspB and corresponding recombinant proteins. (B) TspB NT (lane 1) and CT (lane 2) derivatives resolved

    Article Snippet: As shown in , the NT protein bound to the IgG2 but not IgG1 or IgG3 human paraprotein standards from EMD Millipore.

    Techniques: Binding Assay, Recombinant

    Fluorescence micrographs of complexes formed by purified recombinant TspB derivatives (indicated at left), purified polyclonal IgG, and plasmid DNA. Fluorescence (light areas) indicates DNA and human IgG staining.

    Journal: Infection and Immunity

    Article Title: Resistance of Neisseria meningitidis to Human Serum Depends on T and B Cell Stimulating Protein B

    doi: 10.1128/IAI.03134-14

    Figure Lengend Snippet: Fluorescence micrographs of complexes formed by purified recombinant TspB derivatives (indicated at left), purified polyclonal IgG, and plasmid DNA. Fluorescence (light areas) indicates DNA and human IgG staining.

    Article Snippet: As shown in , the NT protein bound to the IgG2 but not IgG1 or IgG3 human paraprotein standards from EMD Millipore.

    Techniques: Fluorescence, Purification, Recombinant, Plasmid Preparation, Staining