goat anti mouse igg  (Millipore)


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    Structured Review

    Millipore goat anti mouse igg
    Six-to-eight-week-old BALB/c mice (n = 6) and SD rats (n = 6) were twice immunized ID or IM with 3, 9, or 15 µg of HA of the split H7N9 influenza vaccine at 2-week intervals, and serum samples were collected at 2, 3, and 4 weeks post-boosting for HI antibody measurements (A, B). Mice and rats were immunized with the H7N9 influenza vaccine at 0 and 14 days, and serum HI antibody titers and <t>IgG</t> subtypes were assayed at 2 and 4 weeks post-boosting (C).
    Goat Anti Mouse Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 75 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg - by Bioz Stars, 2020-02
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    Images

    1) Product Images from "Intradermal delivery of a fractional dose of influenza H7N9 split vaccine elicits protective immunity in mice and rats"

    Article Title: Intradermal delivery of a fractional dose of influenza H7N9 split vaccine elicits protective immunity in mice and rats

    Journal: Human Vaccines & Immunotherapeutics

    doi: 10.1080/21645515.2017.1423156

    Six-to-eight-week-old BALB/c mice (n = 6) and SD rats (n = 6) were twice immunized ID or IM with 3, 9, or 15 µg of HA of the split H7N9 influenza vaccine at 2-week intervals, and serum samples were collected at 2, 3, and 4 weeks post-boosting for HI antibody measurements (A, B). Mice and rats were immunized with the H7N9 influenza vaccine at 0 and 14 days, and serum HI antibody titers and IgG subtypes were assayed at 2 and 4 weeks post-boosting (C).
    Figure Legend Snippet: Six-to-eight-week-old BALB/c mice (n = 6) and SD rats (n = 6) were twice immunized ID or IM with 3, 9, or 15 µg of HA of the split H7N9 influenza vaccine at 2-week intervals, and serum samples were collected at 2, 3, and 4 weeks post-boosting for HI antibody measurements (A, B). Mice and rats were immunized with the H7N9 influenza vaccine at 0 and 14 days, and serum HI antibody titers and IgG subtypes were assayed at 2 and 4 weeks post-boosting (C).

    Techniques Used: Mouse Assay

    2) Product Images from "Interleukin-22 as a molecular adjuvant facilitates IL-17-producing CD8+ T cell responses against a HBV DNA vaccine in mice"

    Article Title: Interleukin-22 as a molecular adjuvant facilitates IL-17-producing CD8+ T cell responses against a HBV DNA vaccine in mice

    Journal: Human Vaccines & Immunotherapeutics

    doi: 10.4161/hv.26047

    Figure 2. IL-22 as a molecular adjuvant failed to enhance anti-HBsAg humoral response. Sera from immunized mice was collected and pooled 7 d after the third immunization and analyzed by ELISA; ( A ) Total IgG was quantified with a commercial IgG standard; ( B ) IgG1 and ( C ) IgG2a isotypes against HBsAg were quantified against dilutions of standard mouse IgG subtypes. Data shown are representative of three independent experiments (ns, p > 0.05).
    Figure Legend Snippet: Figure 2. IL-22 as a molecular adjuvant failed to enhance anti-HBsAg humoral response. Sera from immunized mice was collected and pooled 7 d after the third immunization and analyzed by ELISA; ( A ) Total IgG was quantified with a commercial IgG standard; ( B ) IgG1 and ( C ) IgG2a isotypes against HBsAg were quantified against dilutions of standard mouse IgG subtypes. Data shown are representative of three independent experiments (ns, p > 0.05).

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    3) Product Images from "Experimental Infection of Mongolian Gerbils with Wild-Type and Mutant Helicobacter pylori Strains"

    Article Title: Experimental Infection of Mongolian Gerbils with Wild-Type and Mutant Helicobacter pylori Strains

    Journal: Infection and Immunity

    doi:

    Variability of the serum antibody response to H. pylori in six representative infected gerbils during the first 12 weeks following challenge with strain G1.1. (A) IgM. (B) IgG. Blood specimens were obtained prechallenge, weekly from weeks 2 to 6, and at 8 and 12 weeks. Antibody levels in serum were determined by ELISA with a sonicate of G1.1 cells as the antigen. The results shown for each gerbil represent the mean of two duplicate determinations. The symbols for each gerbil are the same in both panels. (C) Mean values for IgG (circles) or IgM (squares) and SEM in 21 animals.
    Figure Legend Snippet: Variability of the serum antibody response to H. pylori in six representative infected gerbils during the first 12 weeks following challenge with strain G1.1. (A) IgM. (B) IgG. Blood specimens were obtained prechallenge, weekly from weeks 2 to 6, and at 8 and 12 weeks. Antibody levels in serum were determined by ELISA with a sonicate of G1.1 cells as the antigen. The results shown for each gerbil represent the mean of two duplicate determinations. The symbols for each gerbil are the same in both panels. (C) Mean values for IgG (circles) or IgM (squares) and SEM in 21 animals.

    Techniques Used: Infection, Enzyme-linked Immunosorbent Assay

    4) Product Images from "Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice"

    Article Title: Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-53135-z

    Immunization with rSsEno with or without PGA conjugation enhanced the antibody response. BALB/c mice were s.c. immunized three times with rSsEno100, PGA + rSsEno100 or PBS as a negative control. Sera collected seven days after the last boost was used to determine antigen-specific IgG ( A ), IgG1 ( B ), IgG2a ( C ), and IgG3 ( D ) titers by ELISA. The results are presented as the mean ± SD of 5 mice from one of three independent experiments, and statistical significance was determined by one-way ANOVA using Tukey’s multiple comparisons test and a 95% confidence interval. *(p
    Figure Legend Snippet: Immunization with rSsEno with or without PGA conjugation enhanced the antibody response. BALB/c mice were s.c. immunized three times with rSsEno100, PGA + rSsEno100 or PBS as a negative control. Sera collected seven days after the last boost was used to determine antigen-specific IgG ( A ), IgG1 ( B ), IgG2a ( C ), and IgG3 ( D ) titers by ELISA. The results are presented as the mean ± SD of 5 mice from one of three independent experiments, and statistical significance was determined by one-way ANOVA using Tukey’s multiple comparisons test and a 95% confidence interval. *(p

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

    5) Product Images from "Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses"

    Article Title: Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses

    Journal: Iranian Journal of Basic Medical Sciences

    doi: 10.22038/IJBMS.2018.29112.7026

    Anti-F1S1 specific antibodies in serum. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgG (a), IgG1 (b), IgG2a (c) and IgG2a/IgG1 ratio (d) were assayed in sera by ELISA. Results depict A450 of sera diluted (1:100) and are expressed as mean + SD of 5 mice in each group (* P
    Figure Legend Snippet: Anti-F1S1 specific antibodies in serum. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgG (a), IgG1 (b), IgG2a (c) and IgG2a/IgG1 ratio (d) were assayed in sera by ELISA. Results depict A450 of sera diluted (1:100) and are expressed as mean + SD of 5 mice in each group (* P

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

    The kinetics of the F1S1-specific serum IgG antibody responses. Sera from 5 mice in each group were pooled and assayed in triplicates. The comparisons were made with adjuvant-treated control groups. (**** P
    Figure Legend Snippet: The kinetics of the F1S1-specific serum IgG antibody responses. Sera from 5 mice in each group were pooled and assayed in triplicates. The comparisons were made with adjuvant-treated control groups. (**** P

    Techniques Used: Mouse Assay

    Specific anti-F1S1 antibody levels in lung homogenates. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgA (a), IgG (b), and IgA to IgG ratio (c) were determined based on the measured secreted antibodies in the lung homogenates by ELISA. Results are expressed as mean + SD of 5 mice in each group (*** P
    Figure Legend Snippet: Specific anti-F1S1 antibody levels in lung homogenates. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgA (a), IgG (b), and IgA to IgG ratio (c) were determined based on the measured secreted antibodies in the lung homogenates by ELISA. Results are expressed as mean + SD of 5 mice in each group (*** P

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

    6) Product Images from "Mechanistic Insight into the TH1-Biased Immune Response to Recombinant Subunit Vaccines Delivered by Probiotic Bacteria-Derived Outer Membrane Vesicles"

    Article Title: Mechanistic Insight into the TH1-Biased Immune Response to Recombinant Subunit Vaccines Delivered by Probiotic Bacteria-Derived Outer Membrane Vesicles

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0112802

    EcN OMV antigen carriers generate robust, T H 1-biased antigen-specific humoral and cellular immunity in a mouse model. a, Vaccination protocol for immunization of BALB/c mice against a model antigen. b,c, Time-evolution (upper panel) and terminal titers (lower panel) of antigen-specific IgG (b) and IgM (c) from BALB/c mice vaccinated (primed) and boosted once with antigen-normalized doses (n = 5, each group). Experimental groups indicated are as follows: mice injected with a mixture of recombinant ClyA-GFP and alum, ClyA-GFP+Alum; with EcN OMVs displaying ClyA-GFP, EcN ClyA-GFP; with EcJ OMVs displaying ClyA-GFP, EcJ ClyA-GFP; with recombinant ClyA in PBS alone, ClyA; with a mixture of ClyA and GFP in PBS, GFP+ClyA; with recombinant GFP in PBS alone, GFP; with EcN OMVs not displaying any recombinant ClyA-antigen fusion, EcN Empty; with EcJ OMVs not displaying any recombinant ClyA-antigen fusion, EcJ Empty; and with PBS alone, PBS. d, Ratio of IgG2a:IgG1 antibodies from terminal total IgG titers in b. e-i, CFSE-measured proliferation index (e) , IFN- γ secretion (f) and expression (g) , IL-4 secretion (h) , and IL-10 secretion (i) from cultured, GFP-restimulated splenic T-cells harvested from end-point subjects used in b-d (n = 5, each group). GFP* data are representative of GFP, ClyA, GFP+ClyA, EcJ Empty and EcN Empty, and PBS control groups. **P
    Figure Legend Snippet: EcN OMV antigen carriers generate robust, T H 1-biased antigen-specific humoral and cellular immunity in a mouse model. a, Vaccination protocol for immunization of BALB/c mice against a model antigen. b,c, Time-evolution (upper panel) and terminal titers (lower panel) of antigen-specific IgG (b) and IgM (c) from BALB/c mice vaccinated (primed) and boosted once with antigen-normalized doses (n = 5, each group). Experimental groups indicated are as follows: mice injected with a mixture of recombinant ClyA-GFP and alum, ClyA-GFP+Alum; with EcN OMVs displaying ClyA-GFP, EcN ClyA-GFP; with EcJ OMVs displaying ClyA-GFP, EcJ ClyA-GFP; with recombinant ClyA in PBS alone, ClyA; with a mixture of ClyA and GFP in PBS, GFP+ClyA; with recombinant GFP in PBS alone, GFP; with EcN OMVs not displaying any recombinant ClyA-antigen fusion, EcN Empty; with EcJ OMVs not displaying any recombinant ClyA-antigen fusion, EcJ Empty; and with PBS alone, PBS. d, Ratio of IgG2a:IgG1 antibodies from terminal total IgG titers in b. e-i, CFSE-measured proliferation index (e) , IFN- γ secretion (f) and expression (g) , IL-4 secretion (h) , and IL-10 secretion (i) from cultured, GFP-restimulated splenic T-cells harvested from end-point subjects used in b-d (n = 5, each group). GFP* data are representative of GFP, ClyA, GFP+ClyA, EcJ Empty and EcN Empty, and PBS control groups. **P

    Techniques Used: Mouse Assay, Injection, Recombinant, Expressing, Cell Culture

    7) Product Images from "B cells establish, but do not maintain, long-lived murine anti-peanut IgE"

    Article Title: B cells establish, but do not maintain, long-lived murine anti-peanut IgE

    Journal: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

    doi: 10.1111/cea.12715

    Total serum IgG, peanut-specific IgG, and total IgE antibody secreting cells in peanut-allergic mice after 18 weeks of treatment with either anti-CD20 or isotype control antibody. (a) Serum total IgG levels after 18 weeks of treatment with anti-CD20 or
    Figure Legend Snippet: Total serum IgG, peanut-specific IgG, and total IgE antibody secreting cells in peanut-allergic mice after 18 weeks of treatment with either anti-CD20 or isotype control antibody. (a) Serum total IgG levels after 18 weeks of treatment with anti-CD20 or

    Techniques Used: Mouse Assay

    8) Product Images from "Expression and Characterization of Yeast Derived Chikungunya Virus Like Particles (CHIK-VLPs) and Its Evaluation as a Potential Vaccine Candidate"

    Article Title: Expression and Characterization of Yeast Derived Chikungunya Virus Like Particles (CHIK-VLPs) and Its Evaluation as a Potential Vaccine Candidate

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0004782

    In vivo virus neutralization activity of mice sera immunized with CHIK-VLPs. (A) Percentage survival of the all the mice groups. CHIKV infected mice showed 100% mortality whereas treated mice (infected with DRDE 07) that received CHIK-VLPs IgG and then infected with CHIKV showed 100% survival rate same as mock infected mice that neither infected with CHIKV nor received specific IgG. However, treated mice (infected with DRDE 06), showed 90% survival. (B) Body weight gain measured on 1–7 day of post infection. Treated mice group (infected with DRDE 07 or DRDE 06) showed significantly higher (***P
    Figure Legend Snippet: In vivo virus neutralization activity of mice sera immunized with CHIK-VLPs. (A) Percentage survival of the all the mice groups. CHIKV infected mice showed 100% mortality whereas treated mice (infected with DRDE 07) that received CHIK-VLPs IgG and then infected with CHIKV showed 100% survival rate same as mock infected mice that neither infected with CHIKV nor received specific IgG. However, treated mice (infected with DRDE 06), showed 90% survival. (B) Body weight gain measured on 1–7 day of post infection. Treated mice group (infected with DRDE 07 or DRDE 06) showed significantly higher (***P

    Techniques Used: In Vivo, Neutralization, Activity Assay, Mouse Assay, Infection

    Measurement of serum IgG isotypes titers in immunized BALB/c mice. Profile of IgG isotypes in sera from immunized animal groups (40 μg, 20 μg and 10 μg CHIK-VLPs). Data represented as mean antibody titers with S.D. of ten Balb/c mice in each group Analysis was done by one way ANOVA, (Fisher LSD) # P
    Figure Legend Snippet: Measurement of serum IgG isotypes titers in immunized BALB/c mice. Profile of IgG isotypes in sera from immunized animal groups (40 μg, 20 μg and 10 μg CHIK-VLPs). Data represented as mean antibody titers with S.D. of ten Balb/c mice in each group Analysis was done by one way ANOVA, (Fisher LSD) # P

    Techniques Used: Mouse Assay

    Measurement of serum IgG antibody titers in Balb/C mice immunized with CHIK-VLPs. Sera collected after first booster 14, 28, 42, 56 and 140 days of post-vaccination from immunized groups with 40 μg, 20 μg and 10 μg CHIK-VLPs and antibody titer was measured by indirect ELISA. Data represented as mean antibody titers with S.D. of ten Balb/c mice in each group. Analysis was done by one way ANOVA (Fisher LSD Method). ****P
    Figure Legend Snippet: Measurement of serum IgG antibody titers in Balb/C mice immunized with CHIK-VLPs. Sera collected after first booster 14, 28, 42, 56 and 140 days of post-vaccination from immunized groups with 40 μg, 20 μg and 10 μg CHIK-VLPs and antibody titer was measured by indirect ELISA. Data represented as mean antibody titers with S.D. of ten Balb/c mice in each group. Analysis was done by one way ANOVA (Fisher LSD Method). ****P

    Techniques Used: Mouse Assay, Indirect ELISA

    9) Product Images from "An efficient system to generate monoclonal antibodies against membrane-associated proteins by immunisation with antigen-expressing mammalian cells"

    Article Title: An efficient system to generate monoclonal antibodies against membrane-associated proteins by immunisation with antigen-expressing mammalian cells

    Journal: BMC Biotechnology

    doi: 10.1186/1472-6750-10-87

    Reactivity of generated monoclonal antibodies with the endogenous P. falciparum proteins . Fluorescence staining (column 2, 3 4) and DIC micrographs (column 1) of P. falciparum parasites. While anti-PFF0620C mAb specifically stained salivary gland sporozoites (line 5), anti-PF14_0325 mAb (line 3) and anti-PFD1130W mAb (line 2) specifically reacted with schizont blood stage parasites. Staining with only the Cy3-labelled rabbit anti-mouse IgG secondary antibody served as negative control (line 1 4). Parasite nuclei were stained with DAPI.
    Figure Legend Snippet: Reactivity of generated monoclonal antibodies with the endogenous P. falciparum proteins . Fluorescence staining (column 2, 3 4) and DIC micrographs (column 1) of P. falciparum parasites. While anti-PFF0620C mAb specifically stained salivary gland sporozoites (line 5), anti-PF14_0325 mAb (line 3) and anti-PFD1130W mAb (line 2) specifically reacted with schizont blood stage parasites. Staining with only the Cy3-labelled rabbit anti-mouse IgG secondary antibody served as negative control (line 1 4). Parasite nuclei were stained with DAPI.

    Techniques Used: Generated, Fluorescence, Staining, Negative Control

    10) Product Images from "A Novel Cellular Protein, p60, Interacting with both Herpes Simplex Virus 1 Regulatory Proteins ICP22 and ICP0 Is Modified in a Cell-Type-Specific Manner and Is Recruited to the Nucleus after Infection"

    Article Title: A Novel Cellular Protein, p60, Interacting with both Herpes Simplex Virus 1 Regulatory Proteins ICP22 and ICP0 Is Modified in a Cell-Type-Specific Manner and Is Recruited to the Nucleus after Infection

    Journal: Journal of Virology

    doi:

    Photomicrographs of uninfected (A and E) and HSV-1(F)-infected (B to D and F to H) HEp-2 cells (A to D) and rabbit skin cells (E to H) reacted with antibodies to p60 and ICP0. Cells were fixed 18 h after infection and processed for immunofluorescence as described in Materials and Methods. (A, B, E, and F) Texas red-labeled anti-rabbit IgG to p60 serum; (C and G) FITC-labeled anti-mouse IgG to the ICP0 antibody; (D and H) overlays of panels B and C and panels F and G, respectively.
    Figure Legend Snippet: Photomicrographs of uninfected (A and E) and HSV-1(F)-infected (B to D and F to H) HEp-2 cells (A to D) and rabbit skin cells (E to H) reacted with antibodies to p60 and ICP0. Cells were fixed 18 h after infection and processed for immunofluorescence as described in Materials and Methods. (A, B, E, and F) Texas red-labeled anti-rabbit IgG to p60 serum; (C and G) FITC-labeled anti-mouse IgG to the ICP0 antibody; (D and H) overlays of panels B and C and panels F and G, respectively.

    Techniques Used: Infection, Immunofluorescence, Labeling

    11) Product Images from "Expression of tetanus toxin Fragment C in tobacco chloroplasts"

    Article Title: Expression of tetanus toxin Fragment C in tobacco chloroplasts

    Journal: Nucleic Acids Research

    doi:

    Response to nasal vaccination with plant-produced TetC. ( A ) Immunization schedule and dosing. ( B ) Serum anti-TetC IgG antibody titer indicates systemic immune response. ( C ) Nasal anti-TetC IgA titer. ( D ) Gut anti-TetC IgA titer. Open symbols are values after 27 days; filled symbols are values after 45 days. The bars are averages of five measurements.
    Figure Legend Snippet: Response to nasal vaccination with plant-produced TetC. ( A ) Immunization schedule and dosing. ( B ) Serum anti-TetC IgG antibody titer indicates systemic immune response. ( C ) Nasal anti-TetC IgA titer. ( D ) Gut anti-TetC IgA titer. Open symbols are values after 27 days; filled symbols are values after 45 days. The bars are averages of five measurements.

    Techniques Used: Produced

    12) Product Images from "Utilization of Complement-Dependent Cytotoxicity To Measure Low Levels of Antibodies: Application to Nonstructural Protein 1 in a Model of Japanese Encephalitis Virus ▿"

    Article Title: Utilization of Complement-Dependent Cytotoxicity To Measure Low Levels of Antibodies: Application to Nonstructural Protein 1 in a Model of Japanese Encephalitis Virus ▿

    Journal:

    doi: 10.1128/CVI.00347-07

    Time course of NS1 antibody levels/titers in mice infected with JEV. Sera were examined by the one-dilution (A) or endpoint (B) method of the CDC assay and ELISA for measuring IgG (C) or IgM (D) antibodies. The sera used in this experiment were collected
    Figure Legend Snippet: Time course of NS1 antibody levels/titers in mice infected with JEV. Sera were examined by the one-dilution (A) or endpoint (B) method of the CDC assay and ELISA for measuring IgG (C) or IgM (D) antibodies. The sera used in this experiment were collected

    Techniques Used: Mouse Assay, Infection, CDC Assay, Enzyme-linked Immunosorbent Assay

    Percentages of specific cell lysis obtained with the day-4 serum or HMS depleted of IgG or IgM as determined by the one-dilution method of the CDC assay. Serum was incubated with anti-mouse IgG or IgM and examined for depletion by a sandwich ELISA for
    Figure Legend Snippet: Percentages of specific cell lysis obtained with the day-4 serum or HMS depleted of IgG or IgM as determined by the one-dilution method of the CDC assay. Serum was incubated with anti-mouse IgG or IgM and examined for depletion by a sandwich ELISA for

    Techniques Used: Lysis, CDC Assay, Incubation, Sandwich ELISA

    13) Product Images from "Protective immunity induced by peptides of AMA1, RON2 and RON4 containing T-and B-cell epitopes via an intranasal route against toxoplasmosis in mice"

    Article Title: Protective immunity induced by peptides of AMA1, RON2 and RON4 containing T-and B-cell epitopes via an intranasal route against toxoplasmosis in mice

    Journal: Parasites & Vectors

    doi: 10.1186/s13071-015-0636-5

    Nasal immunization induces antigen-specific IgA and IgG responses in sera. Serum samples were collected and diluted 1:300 to analyze the (A) IgA, (B) total IgG, (C) IgG1 and IgG2a contents by ELISA 14 days after the last immunization. The results are expressed as the means of the OD 492 ± SD (n = 6) and from the two - time determinations. a vs. b, P
    Figure Legend Snippet: Nasal immunization induces antigen-specific IgA and IgG responses in sera. Serum samples were collected and diluted 1:300 to analyze the (A) IgA, (B) total IgG, (C) IgG1 and IgG2a contents by ELISA 14 days after the last immunization. The results are expressed as the means of the OD 492 ± SD (n = 6) and from the two - time determinations. a vs. b, P

    Techniques Used: Enzyme-linked Immunosorbent Assay

    14) Product Images from "Tumor Necrosis Factor Alpha Is a Key Mediator in the Regulation of Experimental Trypanosoma brucei Infections"

    Article Title: Tumor Necrosis Factor Alpha Is a Key Mediator in the Regulation of Experimental Trypanosoma brucei Infections

    Journal: Infection and Immunity

    doi:

    Development of an antiflagellar pocket immune response during the experimental infection with T. brucei AnTat 1.1. Antibody titers were determined with the serum of both infected wild-type and TNF-α −/− C57BL/6 mice. Preimmune serum was used to determine aspecific binding. Serum samples were analyzed in triplicate (mean ± standard deviation) at day 6 (A), day 7 (B), day 14 (C), and day 35 (D), and serum antibody titers were checked for the wild-type IgM (●) and IgG (■) responses and the TNF-α −/− IgM (○) and IgG (□) responses.
    Figure Legend Snippet: Development of an antiflagellar pocket immune response during the experimental infection with T. brucei AnTat 1.1. Antibody titers were determined with the serum of both infected wild-type and TNF-α −/− C57BL/6 mice. Preimmune serum was used to determine aspecific binding. Serum samples were analyzed in triplicate (mean ± standard deviation) at day 6 (A), day 7 (B), day 14 (C), and day 35 (D), and serum antibody titers were checked for the wild-type IgM (●) and IgG (■) responses and the TNF-α −/− IgM (○) and IgG (□) responses.

    Techniques Used: Infection, Mouse Assay, Binding Assay, Standard Deviation

    15) Product Images from "Baculovirus infection induces disruption of the nuclear lamina"

    Article Title: Baculovirus infection induces disruption of the nuclear lamina

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-08437-5

    Subcellular localization of GFP-lamin B in Sf9-L cells. ( A ) Immunofluorescence microscopy analysis. Sf9-L cells were mock-infected or infected with vAc76:FLAG at an MOI of 5. At the designated time points p.i., the cells were fixed, permeabilized, stained with a mouse monoclonal anti-FLAG antibody, and visualized with donkey anti-mouse IgG conjugated to Alexa Fluor 555 (red) as the secondary antibody to detect Ac76. GFP-lamin B was detected based on GFP autofluorescence (GFP-Auto, green). Hoechst 33342 was used to identify nuclei and DNA-rich regions (blue). The red rectangles indicate the diffuse localization of a portion of the nuclear rim GFP-lamin B, and the red triangles show the distribution of GFP-lamin B at the periphery of the nucleus. ( B ) Higher magnification image of the boxed region in ( Aa ). ( C ) Live confocal microscopy images of a virus-infected Sf9-L cell between 17 h 50 min and 18 h 30 min p.i. A portion of the nuclear rim GFP-lamin B showed diffuse localization (red arrow) and then recovered (white arrow).
    Figure Legend Snippet: Subcellular localization of GFP-lamin B in Sf9-L cells. ( A ) Immunofluorescence microscopy analysis. Sf9-L cells were mock-infected or infected with vAc76:FLAG at an MOI of 5. At the designated time points p.i., the cells were fixed, permeabilized, stained with a mouse monoclonal anti-FLAG antibody, and visualized with donkey anti-mouse IgG conjugated to Alexa Fluor 555 (red) as the secondary antibody to detect Ac76. GFP-lamin B was detected based on GFP autofluorescence (GFP-Auto, green). Hoechst 33342 was used to identify nuclei and DNA-rich regions (blue). The red rectangles indicate the diffuse localization of a portion of the nuclear rim GFP-lamin B, and the red triangles show the distribution of GFP-lamin B at the periphery of the nucleus. ( B ) Higher magnification image of the boxed region in ( Aa ). ( C ) Live confocal microscopy images of a virus-infected Sf9-L cell between 17 h 50 min and 18 h 30 min p.i. A portion of the nuclear rim GFP-lamin B showed diffuse localization (red arrow) and then recovered (white arrow).

    Techniques Used: Immunofluorescence, Microscopy, Infection, Staining, Confocal Microscopy

    Generation of the Sf9-L clonal cell line. ( A ) Subcellular localization of lamin B in Sf9-L cells. The cells were fixed, permeabilized, stained with the mouse monoclonal antibody ADL67, and visualized with donkey anti-mouse IgG conjugated to Alexa Fluor 555 (red) as the secondary antibody to detect both native lamin B and chimeric GFP-lamin B. Chimeric GFP-lamin B was detected by visualizing GFP autofluorescence (GFP-Auto, green). Hoechst 33342 was used to identify the nuclei and DNA-rich regions (blue). ( B ) Time course analysis of total GFP-lamin B. Sf9-L cells were mock-infected or infected with vAcWT at an MOI of 10. At the indicated time points, the cells were collected, resolved by SDS-10% PAGE, and subjected to Western blotting with a mouse monoclonal antibody against GFP and an anti-actin antibody as a loading control. Mi, mock-infected Sf9-L cells. ( C ) Viral replication analysis in Sf9-L and Sf9 cells. Sf9-L and Sf9 cells were infected with vAcWT-mCh at an MOI of 5, and the supernatants were harvested at the selected time points. The titers were determined using TCID 50 assays. Each data point represents the average of three independent infections. The error bars represent the standard deviations.
    Figure Legend Snippet: Generation of the Sf9-L clonal cell line. ( A ) Subcellular localization of lamin B in Sf9-L cells. The cells were fixed, permeabilized, stained with the mouse monoclonal antibody ADL67, and visualized with donkey anti-mouse IgG conjugated to Alexa Fluor 555 (red) as the secondary antibody to detect both native lamin B and chimeric GFP-lamin B. Chimeric GFP-lamin B was detected by visualizing GFP autofluorescence (GFP-Auto, green). Hoechst 33342 was used to identify the nuclei and DNA-rich regions (blue). ( B ) Time course analysis of total GFP-lamin B. Sf9-L cells were mock-infected or infected with vAcWT at an MOI of 10. At the indicated time points, the cells were collected, resolved by SDS-10% PAGE, and subjected to Western blotting with a mouse monoclonal antibody against GFP and an anti-actin antibody as a loading control. Mi, mock-infected Sf9-L cells. ( C ) Viral replication analysis in Sf9-L and Sf9 cells. Sf9-L and Sf9 cells were infected with vAcWT-mCh at an MOI of 5, and the supernatants were harvested at the selected time points. The titers were determined using TCID 50 assays. Each data point represents the average of three independent infections. The error bars represent the standard deviations.

    Techniques Used: Staining, Infection, Polyacrylamide Gel Electrophoresis, Western Blot

    16) Product Images from "SAG4 DNA and Peptide Vaccination Provides Partial Protection against T. gondii Infection in BALB/c Mice"

    Article Title: SAG4 DNA and Peptide Vaccination Provides Partial Protection against T. gondii Infection in BALB/c Mice

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2017.01733

    Determination of specific anti- T. gondii IgG1 and IgG2a in immunized mice. The sera of 20 mice of each group were collected from 2 weeks after the final immunization and the IgG subtypes levels were analyzed by ELISA. T. gondii -specific IgG1 and IgG2a were used in the detection. Results are expressed as means of the OD490 ± SD statistically significant differences ( p
    Figure Legend Snippet: Determination of specific anti- T. gondii IgG1 and IgG2a in immunized mice. The sera of 20 mice of each group were collected from 2 weeks after the final immunization and the IgG subtypes levels were analyzed by ELISA. T. gondii -specific IgG1 and IgG2a were used in the detection. Results are expressed as means of the OD490 ± SD statistically significant differences ( p

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Measurement of specific IgG antibodies in immunized sera of mice. Twenty blood samples of each group were collected from caudal vein on days 13, 27, and 41. Results are expressed as means of the OD490 of the samples. T. gondii -specific IgG was used in the study. All the samples were run three times. ∗ Compared with PBS or pEGFP-C1, p
    Figure Legend Snippet: Measurement of specific IgG antibodies in immunized sera of mice. Twenty blood samples of each group were collected from caudal vein on days 13, 27, and 41. Results are expressed as means of the OD490 of the samples. T. gondii -specific IgG was used in the study. All the samples were run three times. ∗ Compared with PBS or pEGFP-C1, p

    Techniques Used: Mouse Assay

    17) Product Images from "Characteristics of Invasive Candidiasis in Gamma Interferon- and Interleukin-4-Deficient Mice: Role of Macrophages in Host Defense against Candida albicans"

    Article Title: Characteristics of Invasive Candidiasis in Gamma Interferon- and Interleukin-4-Deficient Mice: Role of Macrophages in Host Defense against Candida albicans

    Journal: Infection and Immunity

    doi:

    (A) C. albicans -specific IgM, IgG, and IgA titers in sera of mice infected i.p. with 10 8 Candida blastoconidia. Pooled sera of at least four mice/group were analyzed for specific antibodies by standard ELISA. The height of each bar represents the mean ± standard error of three experiments performed in duplicate. (B) C. albicans -specific IgG1, IgG3, IgG2a, and IgG2b titers in sera of mice infected i.p. with 10 8 Candida blastoconidia. Pooled serum samples of at least four mice per group were analyzed for specific antibodies by standard ELISA. The height of each bar represents the mean ± standard error of three experiments performed in duplicate.
    Figure Legend Snippet: (A) C. albicans -specific IgM, IgG, and IgA titers in sera of mice infected i.p. with 10 8 Candida blastoconidia. Pooled sera of at least four mice/group were analyzed for specific antibodies by standard ELISA. The height of each bar represents the mean ± standard error of three experiments performed in duplicate. (B) C. albicans -specific IgG1, IgG3, IgG2a, and IgG2b titers in sera of mice infected i.p. with 10 8 Candida blastoconidia. Pooled serum samples of at least four mice per group were analyzed for specific antibodies by standard ELISA. The height of each bar represents the mean ± standard error of three experiments performed in duplicate.

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

    18) Product Images from "Actin-Binding Protein 1 Regulates B Cell Receptor-Mediated Antigen Processing and Presentation in Response to B Cell Receptor Activation"

    Article Title: Actin-Binding Protein 1 Regulates B Cell Receptor-Mediated Antigen Processing and Presentation in Response to B Cell Receptor Activation

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

    doi:

    BCR activation induces the redistribution of Abp1 to the plasma membrane. A and C , A20 ( A ) and splenic B cells ( C ) were treated (+XL) or untreated (−XL) with either goat anti-mouse IgG ( A, a, f , and k ) or AF488 goat anti-mouse IgG + M ( A, b–e, g–j , and l–o , and C ) at 4°C and chased at 37°C for indicated times. After fixation and permeabilization, cells were labeled with anti-Abp1 Ab for the endogenously expressed Abp1 ( A, a, f , and k , and C ) or anti- myc Ab for myc -Abp1 ( A, b–e, g–j , and l–o ). Cells were analyzed using a confocal fluorescence microscope. Shown are representative images from three independent experiments. Bar, 5 µm. B , The redistribution of endogenous Abp1 and myc -Abp1 from the cytoplasm to the cell surface after BCR cross-linking for 2 min was quantified by visual inspection. Over 100 cells from five randomly selected fields of each experiment were analyzed, and the numbers of cells showing Abp1 concentrated on the cell surface were plotted as percentages of the total number of cells inspected. Shown are averages (±SD) from three independent experiments. D , The correlation coefficients of endogenous Abp1 with the BCR in splenic B cells before and after BCR cross-linking was determined using the LSM510 software. Shown are the data generated from > 100 cells of two independent experiments. , Indicate the mean. **, p
    Figure Legend Snippet: BCR activation induces the redistribution of Abp1 to the plasma membrane. A and C , A20 ( A ) and splenic B cells ( C ) were treated (+XL) or untreated (−XL) with either goat anti-mouse IgG ( A, a, f , and k ) or AF488 goat anti-mouse IgG + M ( A, b–e, g–j , and l–o , and C ) at 4°C and chased at 37°C for indicated times. After fixation and permeabilization, cells were labeled with anti-Abp1 Ab for the endogenously expressed Abp1 ( A, a, f , and k , and C ) or anti- myc Ab for myc -Abp1 ( A, b–e, g–j , and l–o ). Cells were analyzed using a confocal fluorescence microscope. Shown are representative images from three independent experiments. Bar, 5 µm. B , The redistribution of endogenous Abp1 and myc -Abp1 from the cytoplasm to the cell surface after BCR cross-linking for 2 min was quantified by visual inspection. Over 100 cells from five randomly selected fields of each experiment were analyzed, and the numbers of cells showing Abp1 concentrated on the cell surface were plotted as percentages of the total number of cells inspected. Shown are averages (±SD) from three independent experiments. D , The correlation coefficients of endogenous Abp1 with the BCR in splenic B cells before and after BCR cross-linking was determined using the LSM510 software. Shown are the data generated from > 100 cells of two independent experiments. , Indicate the mean. **, p

    Techniques Used: Activation Assay, Labeling, Fluorescence, Microscopy, Software, Generated

    Interaction of Abp1 with dynamin 2. A , A20 cells were incubated with (+XL) and without (−XL) goat anti-mouse IgG for 2 min and then fixed, permeabilized, and labeled with anti-Abp1 and anti-dynamin 2 Abs and corresponding secondary Abs. The cells were cotransfected using a confocal fluorescence microscope. Shown are representative images from three independent experiments. Bar, 5 µm. B , A20 cells were cotransfected with myc -Abp1 and GFP-dynamin 2 (GFP-Dyn) ( a–l ) or GFP-dynamin 2 with PRD deletion (GFP-ΔPRD) ( m–x ). The cells were incubated with ( e–l, q–x ) and without (−XL) ( a–d, m–p ) goat anti-mouse IgG for indicated times, and then fixed, permeabilized, and labeled with Cy3 anti- myc Ab to label myc -Abp1. The cells were analyzed by confocal fluorescence microscopy. Shown are representative images from three independent experiments. Bar, 5 µm. C , Shown are the correlation coefficients between myc -Abp1 and GFP-Dyn or GFP-ΔPRD in the cell surface area of > 30 cells from three independent experiments. , Represent the mean correlation coefficient. **, p
    Figure Legend Snippet: Interaction of Abp1 with dynamin 2. A , A20 cells were incubated with (+XL) and without (−XL) goat anti-mouse IgG for 2 min and then fixed, permeabilized, and labeled with anti-Abp1 and anti-dynamin 2 Abs and corresponding secondary Abs. The cells were cotransfected using a confocal fluorescence microscope. Shown are representative images from three independent experiments. Bar, 5 µm. B , A20 cells were cotransfected with myc -Abp1 and GFP-dynamin 2 (GFP-Dyn) ( a–l ) or GFP-dynamin 2 with PRD deletion (GFP-ΔPRD) ( m–x ). The cells were incubated with ( e–l, q–x ) and without (−XL) ( a–d, m–p ) goat anti-mouse IgG for indicated times, and then fixed, permeabilized, and labeled with Cy3 anti- myc Ab to label myc -Abp1. The cells were analyzed by confocal fluorescence microscopy. Shown are representative images from three independent experiments. Bar, 5 µm. C , Shown are the correlation coefficients between myc -Abp1 and GFP-Dyn or GFP-ΔPRD in the cell surface area of > 30 cells from three independent experiments. , Represent the mean correlation coefficient. **, p

    Techniques Used: Incubation, Labeling, Fluorescence, Microscopy

    19) Product Images from "Rapid Generation of Human-Like Neutralizing Monoclonal Antibodies in Urgent Preparedness for Influenza Pandemics and Virulent Infectious Diseases"

    Article Title: Rapid Generation of Human-Like Neutralizing Monoclonal Antibodies in Urgent Preparedness for Influenza Pandemics and Virulent Infectious Diseases

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0066276

    Analysis of lungs from mice treated with mAb 4E6 and 1H10 after infection with H5N1 virus A/Vietnam/1194/04. (A) Viral copy number in the lungs at day 6 post-infection was determined using Q-PCR. The results are expressed as the mean ± SEM. (B) Virus titer in the lungs at day 6 post-infection was measured using a plaque formation assay in MDCK cells. The results are expressed as the mean ± SEM. (C) Histopathology in pulmonary tissue (40x magnification). 1) Mouse treated with mAb 4E6 at 24 hours post-infection. 2) Mouse treated with mAb 1H10 at 24 hours post-infection. 3) Mice injected with an irrelevant IgG at 24 hours post-infection were used as the control. HE: H E staining. IHC: immunohistochemical staining using a mAb against influenza nucleoprotein.
    Figure Legend Snippet: Analysis of lungs from mice treated with mAb 4E6 and 1H10 after infection with H5N1 virus A/Vietnam/1194/04. (A) Viral copy number in the lungs at day 6 post-infection was determined using Q-PCR. The results are expressed as the mean ± SEM. (B) Virus titer in the lungs at day 6 post-infection was measured using a plaque formation assay in MDCK cells. The results are expressed as the mean ± SEM. (C) Histopathology in pulmonary tissue (40x magnification). 1) Mouse treated with mAb 4E6 at 24 hours post-infection. 2) Mouse treated with mAb 1H10 at 24 hours post-infection. 3) Mice injected with an irrelevant IgG at 24 hours post-infection were used as the control. HE: H E staining. IHC: immunohistochemical staining using a mAb against influenza nucleoprotein.

    Techniques Used: Mouse Assay, Infection, Polymerase Chain Reaction, Plaque Formation Assay, Histopathology, Injection, Staining, Immunohistochemistry

    Therapeutic efficacy of mAb 4E6 and 1H10 in a mouse model of H5N1 virus A/Vietnam/1194/04 infection. (A) The Kaplan-Meier survival curve. BALB/c mice (n=7 per group in two separate experiments) were infected intranasally with 10 LD 50 of A/Vietnam/1194/04 and at 24, 48, or 72 hours, were treated with a single intraperitoneal injection of mAb 4E6 or 1H10 at 10 mg/kg body weight. The control group received IgG at 10 mg/kg body weight at 24 hours post-infection. (B) Percentage weight change in mice treated with mAb 4E6 or 1H10 at different time points post-infection. The mice (n=7) were infected via intranasal inoculation with 10 LD50 of A/Vietnam/1203/4, followed by a single intraperitoneal injection of mAbs at 24, 48, or 72 hours post-infection, and their weights were monitored for 14 days. The mice injected with an irrelevant IgG at 24 hours post-infection were used as the control group.
    Figure Legend Snippet: Therapeutic efficacy of mAb 4E6 and 1H10 in a mouse model of H5N1 virus A/Vietnam/1194/04 infection. (A) The Kaplan-Meier survival curve. BALB/c mice (n=7 per group in two separate experiments) were infected intranasally with 10 LD 50 of A/Vietnam/1194/04 and at 24, 48, or 72 hours, were treated with a single intraperitoneal injection of mAb 4E6 or 1H10 at 10 mg/kg body weight. The control group received IgG at 10 mg/kg body weight at 24 hours post-infection. (B) Percentage weight change in mice treated with mAb 4E6 or 1H10 at different time points post-infection. The mice (n=7) were infected via intranasal inoculation with 10 LD50 of A/Vietnam/1203/4, followed by a single intraperitoneal injection of mAbs at 24, 48, or 72 hours post-infection, and their weights were monitored for 14 days. The mice injected with an irrelevant IgG at 24 hours post-infection were used as the control group.

    Techniques Used: Infection, Mouse Assay, Injection

    20) Product Images from "Chimeric Bivalent Virus-Like Particle Vaccine for H5N1 HPAI and ND Confers Protection against a Lethal Challenge in Chickens and Allows a Strategy of Differentiating Infected from Vaccinated Animals (DIVA)"

    Article Title: Chimeric Bivalent Virus-Like Particle Vaccine for H5N1 HPAI and ND Confers Protection against a Lethal Challenge in Chickens and Allows a Strategy of Differentiating Infected from Vaccinated Animals (DIVA)

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0162946

    Analysis of concentrated chimeric VLP: (A) Western blot identification of the HA protein using anti-H5 monoclonal antibody. M, standard molecular size marker (EBM-1032, Elpisbio), HA, chimeric VLPs; (B) Western blot identification of the F/HA chimeric protein using anti-NDV F monoclonal antibody; (C) Western blot identification of the M1 protein using anti-H5N1 anti-serum; (D) TEM identification of chimeric VLP with scale bar; (E) immunogold-labelled chimeric VLP: VLP were probed with anti-H5 monoclonal antibody, counterstained with 10nm gold spheres coupled to anti-mouse IgG and anti-NDV anti-serum, counterstained with 25nm gold spheres coupled to anti-chicken IgG.
    Figure Legend Snippet: Analysis of concentrated chimeric VLP: (A) Western blot identification of the HA protein using anti-H5 monoclonal antibody. M, standard molecular size marker (EBM-1032, Elpisbio), HA, chimeric VLPs; (B) Western blot identification of the F/HA chimeric protein using anti-NDV F monoclonal antibody; (C) Western blot identification of the M1 protein using anti-H5N1 anti-serum; (D) TEM identification of chimeric VLP with scale bar; (E) immunogold-labelled chimeric VLP: VLP were probed with anti-H5 monoclonal antibody, counterstained with 10nm gold spheres coupled to anti-mouse IgG and anti-NDV anti-serum, counterstained with 25nm gold spheres coupled to anti-chicken IgG.

    Techniques Used: Western Blot, Marker, Transmission Electron Microscopy

    21) Product Images from "Modulation of T cell proliferation and cytokine response by Plumbagin, extracted from Plumbago zeylanica in collagen induced arthritis"

    Article Title: Modulation of T cell proliferation and cytokine response by Plumbagin, extracted from Plumbago zeylanica in collagen induced arthritis

    Journal: BMC Complementary and Alternative Medicine

    doi: 10.1186/1472-6882-11-114

    Effect of Plumbagin on collagen type II induced arthritis . Arthritis was induced in two groups (5 per each group) of mice by injecting collagen type II (CII) along with complete Freund's adjuvant (CII+CFA). Control group with CFA alone. Booster dose was given on day 21. One arthritic group was treated with Plumbagin (PZE-6) another treated with olive oil alone. Retroorbital sera were collected at 3, 5, 7 week intervals for the estimation of anti-type II collagen antibody (IgG) by ELISA. Bars represent means±SEM from n = 5. p-value≤0.05 was significant.
    Figure Legend Snippet: Effect of Plumbagin on collagen type II induced arthritis . Arthritis was induced in two groups (5 per each group) of mice by injecting collagen type II (CII) along with complete Freund's adjuvant (CII+CFA). Control group with CFA alone. Booster dose was given on day 21. One arthritic group was treated with Plumbagin (PZE-6) another treated with olive oil alone. Retroorbital sera were collected at 3, 5, 7 week intervals for the estimation of anti-type II collagen antibody (IgG) by ELISA. Bars represent means±SEM from n = 5. p-value≤0.05 was significant.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    22) Product Images from "Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses"

    Article Title: Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses

    Journal: Iranian Journal of Basic Medical Sciences

    doi: 10.22038/IJBMS.2018.29112.7026

    Anti-F1S1 specific antibodies in serum. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgG (a), IgG1 (b), IgG2a (c) and IgG2a/IgG1 ratio (d) were assayed in sera by ELISA. Results depict A450 of sera diluted (1:100) and are expressed as mean + SD of 5 mice in each group (* P
    Figure Legend Snippet: Anti-F1S1 specific antibodies in serum. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgG (a), IgG1 (b), IgG2a (c) and IgG2a/IgG1 ratio (d) were assayed in sera by ELISA. Results depict A450 of sera diluted (1:100) and are expressed as mean + SD of 5 mice in each group (* P

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

    The kinetics of the F1S1-specific serum IgG antibody responses. Sera from 5 mice in each group were pooled and assayed in triplicates. The comparisons were made with adjuvant-treated control groups. (**** P
    Figure Legend Snippet: The kinetics of the F1S1-specific serum IgG antibody responses. Sera from 5 mice in each group were pooled and assayed in triplicates. The comparisons were made with adjuvant-treated control groups. (**** P

    Techniques Used: Mouse Assay

    Specific anti-F1S1 antibody levels in lung homogenates. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgA (a), IgG (b), and IgA to IgG ratio (c) were determined based on the measured secreted antibodies in the lung homogenates by ELISA. Results are expressed as mean + SD of 5 mice in each group (*** P
    Figure Legend Snippet: Specific anti-F1S1 antibody levels in lung homogenates. Two weeks after the final immunization of BALB/c mice with recombinant F1S1 protein (via SC or IN routes) or commercial DTaP vaccine (via SC route), anti- F1S1 IgA (a), IgG (b), and IgA to IgG ratio (c) were determined based on the measured secreted antibodies in the lung homogenates by ELISA. Results are expressed as mean + SD of 5 mice in each group (*** P

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

    23) Product Images from "Endogenous presenilin 1 redistributes to the surface of lamellipodia upon adhesion of Jurkat cells to a collagen matrix"

    Article Title: Endogenous presenilin 1 redistributes to the surface of lamellipodia upon adhesion of Jurkat cells to a collagen matrix

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    Scanning immunoelectron micrographs of Jurkat cells adherent to a collagen matrix. After overnight adhesion to collagen, cells were incubated with mAb to TCR ( A ), CD44 ( B ), and PS1-N ( C – G ) and with affinity-purified rabbit polyclonal antibody to transthyretin ( H ). Bound antibodies were visualized with 5-nm gold-labeled antibody to mouse IgM (TCR), mouse IgG (CD44), and rabbit IgG to PS1-N and transthyretin followed by silver enhancing. (Bar = 11.258 μm; ×3,500.)
    Figure Legend Snippet: Scanning immunoelectron micrographs of Jurkat cells adherent to a collagen matrix. After overnight adhesion to collagen, cells were incubated with mAb to TCR ( A ), CD44 ( B ), and PS1-N ( C – G ) and with affinity-purified rabbit polyclonal antibody to transthyretin ( H ). Bound antibodies were visualized with 5-nm gold-labeled antibody to mouse IgM (TCR), mouse IgG (CD44), and rabbit IgG to PS1-N and transthyretin followed by silver enhancing. (Bar = 11.258 μm; ×3,500.)

    Techniques Used: Incubation, Affinity Purification, Labeling

    24) Product Images from "Insights into the Internalization and Retrograde Trafficking of Dengue 2 Virus in BHK-21 Cells"

    Article Title: Insights into the Internalization and Retrograde Trafficking of Dengue 2 Virus in BHK-21 Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0025229

    DENV-2 E protein associates with dynein during retrograde trafficking. BHK-21 cells infected with DENV-2 at 1 MOI were fixed at different time points p.i. DENV-2 E protein was stained using mouse anti-E MAb followed by rabbit anti-mouse IgG TRITC, saturated with goat anti-mouse IgG and then stained for dynein using mouse anti-dynein MAb followed by donkey anti-mouse IgG Alexa 488. A ] The split images and the intensity profiles showing the colocalization are displayed for each of the time points; 4 h, 12 h, 24 h, 36 h, 48 h, 60 h and 72 h. B ] Mock infected control cells stained for DENV-2 E protein and dynein. C ] The 3D reconstruction of the z stacked deconvolved images showing the beginnings of association of E-dynein at 4 h, followed by D ] strong colocalization at 48 h and then E ] dissociation at 72 h. F ] Co-immunoprecipitation of DENV-2 E-dynein complex. Lysates of DENV-2 infected or uninfected BHK-21 cells were co-immunoprecipitated with anti-DENV IgG antibody and the blot was probed with anti-dynein antibody followed by goat anti-mouse HRP. Lane 1 - DENV-2 infected. BHK-21 cell lysate, Lane 2 - Immunoprecipitated DENV-2 infected BHK-21 cells, Lane 3 -Immunoprecipitated uninfected BHK-21 cells.
    Figure Legend Snippet: DENV-2 E protein associates with dynein during retrograde trafficking. BHK-21 cells infected with DENV-2 at 1 MOI were fixed at different time points p.i. DENV-2 E protein was stained using mouse anti-E MAb followed by rabbit anti-mouse IgG TRITC, saturated with goat anti-mouse IgG and then stained for dynein using mouse anti-dynein MAb followed by donkey anti-mouse IgG Alexa 488. A ] The split images and the intensity profiles showing the colocalization are displayed for each of the time points; 4 h, 12 h, 24 h, 36 h, 48 h, 60 h and 72 h. B ] Mock infected control cells stained for DENV-2 E protein and dynein. C ] The 3D reconstruction of the z stacked deconvolved images showing the beginnings of association of E-dynein at 4 h, followed by D ] strong colocalization at 48 h and then E ] dissociation at 72 h. F ] Co-immunoprecipitation of DENV-2 E-dynein complex. Lysates of DENV-2 infected or uninfected BHK-21 cells were co-immunoprecipitated with anti-DENV IgG antibody and the blot was probed with anti-dynein antibody followed by goat anti-mouse HRP. Lane 1 - DENV-2 infected. BHK-21 cell lysate, Lane 2 - Immunoprecipitated DENV-2 infected BHK-21 cells, Lane 3 -Immunoprecipitated uninfected BHK-21 cells.

    Techniques Used: Infection, Staining, Immunoprecipitation

    25) Product Images from "Cloning approach and functional analysis of anti-intimin single-chain variable fragment (scFv)"

    Article Title: Cloning approach and functional analysis of anti-intimin single-chain variable fragment (scFv)

    Journal: BMC Research Notes

    doi: 10.1186/1756-0500-4-30

    Functional assay of scFv-intimin by immunofluorescence . A . EPEC E2348/69 strain permeabilized with 4% Triton X-100 followed by incubation with 4.4 μg of anti-intimin scFv. The reaction was developed using anti-His antibody and anti-mouse IgG conjugated to FITC. B . E. coli K12 JM109 (negative control) assayed as described for A .
    Figure Legend Snippet: Functional assay of scFv-intimin by immunofluorescence . A . EPEC E2348/69 strain permeabilized with 4% Triton X-100 followed by incubation with 4.4 μg of anti-intimin scFv. The reaction was developed using anti-His antibody and anti-mouse IgG conjugated to FITC. B . E. coli K12 JM109 (negative control) assayed as described for A .

    Techniques Used: Functional Assay, Immunofluorescence, Incubation, Negative Control

    26) Product Images from "Induction of Influenza-Specific Mucosal Immunity by an Attenuated Recombinant Sendai Virus"

    Article Title: Induction of Influenza-Specific Mucosal Immunity by an Attenuated Recombinant Sendai Virus

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0018780

    GP42-H1 vector induces expression of HA in infected cells in vitro . A) CV-1 cells were cultured on glass slides and infected with the indicated viruses (MOI = 10). At 20 hours post-infection, the infected cells were fixed in acetone and evaluated for HA expression by staining with mouse PR/8 anti-sera followed by TRITC conjugated anti-mouse IgG. Slides were mounted with Prolong-Antifade containing DAPI stain and visualized by immunofluorescence microscopy. B) PR/8, GP42-GFP, GP42-H1 or mock infected CV-1 cells were stained with the mouse PR/8 anti-sera. PR/8 infected cell were also stained with pre-immune sera (grey filled histogram) as a control for binding of nonspecific serum proteins. Surface bound mouse antibodies were detected with anti-mouse IgG-PE and analyzed by flow cytometry. C) Eggs were inoculated with PR/8 virus (P), GP42-GFP (G) or GP42-H1 (H) at 1 HA unit per egg. Sucrose purified recombinant viruses and allantoic fluid from PR/8 infected eggs were resolved on SDS-PAGE. SDS-PAGE gel was stained with gel-code blue (left), anti-GFP western blot (middle) and anti-PR8 western blot (right).
    Figure Legend Snippet: GP42-H1 vector induces expression of HA in infected cells in vitro . A) CV-1 cells were cultured on glass slides and infected with the indicated viruses (MOI = 10). At 20 hours post-infection, the infected cells were fixed in acetone and evaluated for HA expression by staining with mouse PR/8 anti-sera followed by TRITC conjugated anti-mouse IgG. Slides were mounted with Prolong-Antifade containing DAPI stain and visualized by immunofluorescence microscopy. B) PR/8, GP42-GFP, GP42-H1 or mock infected CV-1 cells were stained with the mouse PR/8 anti-sera. PR/8 infected cell were also stained with pre-immune sera (grey filled histogram) as a control for binding of nonspecific serum proteins. Surface bound mouse antibodies were detected with anti-mouse IgG-PE and analyzed by flow cytometry. C) Eggs were inoculated with PR/8 virus (P), GP42-GFP (G) or GP42-H1 (H) at 1 HA unit per egg. Sucrose purified recombinant viruses and allantoic fluid from PR/8 infected eggs were resolved on SDS-PAGE. SDS-PAGE gel was stained with gel-code blue (left), anti-GFP western blot (middle) and anti-PR8 western blot (right).

    Techniques Used: Plasmid Preparation, Expressing, Infection, In Vitro, Cell Culture, Staining, Immunofluorescence, Microscopy, Binding Assay, Flow Cytometry, Cytometry, Purification, Recombinant, SDS Page, Western Blot

    GP42-H1 induces HA-specific antibodies in sera. Mice were administered AF, 1LD 50 PR/8 or 10 3 pfu of either GP42-GFP (vector control) or GP42-H1. Blood was collected 14 days after infection and sera were assayed by ELISA for A) PR/8-specific IgG1 B) PR/8-specific IgG2b and C) PR/8-specific IgG2c antibodies. Student's T tests were performed between each group (n = 6 for all groups except the PR/8 group where n = 12). The P values between each group are indicated. Experiments were repeated 2 times with similar results.
    Figure Legend Snippet: GP42-H1 induces HA-specific antibodies in sera. Mice were administered AF, 1LD 50 PR/8 or 10 3 pfu of either GP42-GFP (vector control) or GP42-H1. Blood was collected 14 days after infection and sera were assayed by ELISA for A) PR/8-specific IgG1 B) PR/8-specific IgG2b and C) PR/8-specific IgG2c antibodies. Student's T tests were performed between each group (n = 6 for all groups except the PR/8 group where n = 12). The P values between each group are indicated. Experiments were repeated 2 times with similar results.

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

    GP42-H1 induces HA-specific antibodies at mucosal sites. A) Mice were infected with 0.25 LD 50 (63 pfu) of PR/8 or 10 3 pfu of either GP42-GFP or GP42-H1. At 20 days post infection, mice (n = 4) were sacrificed and BAL fluid was collected and assayed for PR/8-specific IgG1 antibodies. B) At 14 days post-infection, fecal pellet suspensions and C) saliva were collected from the mice (n = 6 for all groups except PR/8 where n = 12) described in figure 4 and assayed for the presence of PR/8 specific IgA antibodies. Shown are representative data from one of three independent experiments.
    Figure Legend Snippet: GP42-H1 induces HA-specific antibodies at mucosal sites. A) Mice were infected with 0.25 LD 50 (63 pfu) of PR/8 or 10 3 pfu of either GP42-GFP or GP42-H1. At 20 days post infection, mice (n = 4) were sacrificed and BAL fluid was collected and assayed for PR/8-specific IgG1 antibodies. B) At 14 days post-infection, fecal pellet suspensions and C) saliva were collected from the mice (n = 6 for all groups except PR/8 where n = 12) described in figure 4 and assayed for the presence of PR/8 specific IgA antibodies. Shown are representative data from one of three independent experiments.

    Techniques Used: Mouse Assay, Infection

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    Bicinchoninic Acid Protein Assay:

    Article Title: Experimental Infection of Mongolian Gerbils with Wild-Type and Mutant Helicobacter pylori Strains
    Article Snippet: Immulon 2 flat-bottom microtiter plates (Dynatech Laboratories, Inc., Chantilly, Va.) were coated with 100 μl of a whole-cell sonicate of H. pylori G1.1, corresponding to 1 μg of protein per well as determined by the bicinchoninic acid protein assay (Pierce, Rockford, Ill.). .. Therefore, bound gerbil IgG antibodies were detected by goat anti-mouse IgG and, by analogy, gerbil IgM was detected by goat anti-mouse IgM antibodies coupled with alkaline phosphatase (Sigma) and p -nitrophenylphosphate as the substrate, using methods described previously ( ).

    Purification:

    Article Title: Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice
    Article Snippet: SDS-PAGE, western blot analysis Samples containing 20 µg of protein S. schenckii ATCC 16345 CWPs and purified rSsEno (5 µg) were resolved on an SDS-PAGE 12% as described by Laemmli . .. After three washes with PBS, the strips were further incubated for 2 h with goat anti-mouse IgG (Sigma-Aldrich) diluted 1:500 or goat anti-feline IgG (Southern Biotech) diluted 1:1000.

    Article Title: Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses
    Article Snippet: In brief, ELISA plates (Greiner, Germany) were coated with 280 ng purified F1S1 and blocked by 1% BSA in PBS (BSA-PBS). .. Goat anti-mouse IgG, rabbit anti-mouse IgG1, rabbit anti-mouse IgG2a, or goat anti-mouse IgA antibodies (Sigma, Germany), all conjugated to horseradish peroxidase, were used as the secondary antibodies.

    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
    Article Snippet: In brief, purified PfCelTOS (20 ng/well) in 0.06 M carbonate-bicarbonate buffer (pH 9.6) was used as an antigen, and MaxiSorp flat-bottom 96-well microplates (Jet Bio-fil Co., Ltd., China) were coated with the antigen and kept at 4°C overnight. .. In order to evaluate the IgG subclass responses to PfCelTOS, the test was performed as described above, and instead of goat anti-mouse IgG-HRP, 100 μl of either goat anti-mouse IgG1, IgG2a, IgG2b, or IgG3 (diluted 1:1,000 in PBS-T buffer) (Sigma-Aldrich) as a secondary antibody was added to each well and incubated at RT for 1 h. After washing, the plates were incubated with a 1:10,000 dilution of anti-goat IgG-HRP (Sigma-Aldrich) at RT for 1 h, and the reaction was then developed by using an enzyme-specific substrate, as mentioned above.

    Article Title: G2 Dendrimer as a Carrier Can Enhance Immune Responses Against HCV-NS3 Protein in BALB/c Mice
    Article Snippet: Briefly, purified rNS3 (3 μg/ml ) was used as a captured molecule to coat 96-well polyvinyl chloride ELISA plates (Nunc, Denmark) for overnight at 4°C . .. IgG antibody subclasses were analyzed as mentioned above using goat anti-mouse IgG1, IgG2a, IgG2b antibodies (Sigma, Aldrich) and rabbit anti-goat IgG-HRP conjugate (Sigma, Aldrich).

    Avidin-Biotin Assay:

    Article Title: Mechanistic Insight into the TH1-Biased Immune Response to Recombinant Subunit Vaccines Delivered by Probiotic Bacteria-Derived Outer Membrane Vesicles
    Article Snippet: Plates were washed six times with PBST, and biotinilated goat anti-mouse IgG, IgM (Sigma), or monoclonal IgG1 or IgG2 (BD Pharmingen) were added to the wells (1 µg/ml) for 1 hour at 37°C. .. Avidin-HRP (1∶1000; Sigma) was then added and incubation continued for 30 min at 37°C.

    Article Title: B cells establish, but do not maintain, long-lived murine anti-peanut IgE
    Article Snippet: Ninety-six-well PVDF membrane ELISPOT plates (Millipore) were coated with CPE, goat anti-mouse IgG (Sigma), or rat anti-mouse IgE (R35-72, BD Biosciences). .. The following Ab were incubated sequentially for one hour at 37°C: biotin goat-anti-mouse IgG (BioLegend) and avidin-peroxidase for IgG ASCs; or sheep anti-mouse IgE (The Binding Site Group, Ltd), rabbit anti-sheep IgG and peroxidase goat anti-rabbit IgG (both from Jackson ImmunoResearch Laboratories Inc.) for IgE ASCs.

    Microscopy:

    Article Title: A Novel Cellular Protein, p60, Interacting with both Herpes Simplex Virus 1 Regulatory Proteins ICP22 and ICP0 Is Modified in a Cell-Type-Specific Manner and Is Recruited to the Nucleus after Infection
    Article Snippet: The cells were rinsed three times with PBS and then reacted for 1 h at room temperature with goat anti-rabbit immunoglobulin G (IgG) conjugated to Texas red (Molecular Probes) and goat anti-mouse IgG conjugated to fluorescein isothiocyanate (FITC; Sigma). .. The slides were examined with a Zeiss confocal fluorescence microscope, and digitized images were acquired with software provided by the manufacturer and printed with a Tektronix 440 phaser printer.

    Mouse Assay:

    Article Title: Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice
    Article Snippet: Sera from naïve mice or sera from cats with no evidence of sporotrichosis (n = 3) were utilized as negative controls. .. After three washes with PBS, the strips were further incubated for 2 h with goat anti-mouse IgG (Sigma-Aldrich) diluted 1:500 or goat anti-feline IgG (Southern Biotech) diluted 1:1000.

    Article Title: Heat Shock Proteins Enriched-Promastigotes of Leishmania major Inducing Th2 Immune Response in BALB/c Mice
    Article Snippet: Ten days after final immunization and two months after infection, mice sera from each group were collected and anti-leishmania antibodies (IgG1, IgG2a, IgG2b and total IgG) were detected by ELISA assay. .. For isotype determination, goat anti-mouse IgG1, IgG2a and IgG2b (Sigma, USA) and HRP-conjugated mouse anti-goat antibody were used according to the manufacture’s instruction.

    Article Title: Intradermal delivery of a fractional dose of influenza H7N9 split vaccine elicits protective immunity in mice and rats
    Article Snippet: Bound antibodies were detected using 1:20,000 dilutions of goat anti-mouse IgG, IgG1, and IgG2a (Sigma, USA) and goat-anti-rat IgG (Sigma), conjugated to horseradish peroxidase. .. Mice that received PBS alone were used as negative controls.

    Article Title: G2 Dendrimer as a Carrier Can Enhance Immune Responses Against HCV-NS3 Protein in BALB/c Mice
    Article Snippet: After washing and blocking steps, wells were probed with optimum dilution of mice sera (1/1000 for total IgG), incubated for 1 hr , washed and further incubated with HRP-conjugated goat anti-mouse IgG (Sigma, Aldrich) as secondary antibody. .. IgG antibody subclasses were analyzed as mentioned above using goat anti-mouse IgG1, IgG2a, IgG2b antibodies (Sigma, Aldrich) and rabbit anti-goat IgG-HRP conjugate (Sigma, Aldrich).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Interleukin-22 as a molecular adjuvant facilitates IL-17-producing CD8+ T cell responses against a HBV DNA vaccine in mice
    Article Snippet: The detection of anti-HBsAg-specific antibodies in the serum was by quantitative ELISA. .. After 5 time washes with PBST, 1:1000 diluted goat anti-mouse IgG, IgG1, or IgG2a conjugated with HPR was added to the wells, and the plates were incubated at 37 °C for 1 h. After five times of wash, 100 μl TMB buffer (Sigma, cat.no.T2885, 10 mg TMB powder dissolved in 200 μl H2 O and 800 μl ethanol) was added to each well.

    Article Title: Mechanistic Insight into the TH1-Biased Immune Response to Recombinant Subunit Vaccines Delivered by Probiotic Bacteria-Derived Outer Membrane Vesicles
    Article Snippet: Paragraph title: ELISA for Serum Antibody Response ... Plates were washed six times with PBST, and biotinilated goat anti-mouse IgG, IgM (Sigma), or monoclonal IgG1 or IgG2 (BD Pharmingen) were added to the wells (1 µg/ml) for 1 hour at 37°C.

    Article Title: Heat Shock Proteins Enriched-Promastigotes of Leishmania major Inducing Th2 Immune Response in BALB/c Mice
    Article Snippet: Ten days after final immunization and two months after infection, mice sera from each group were collected and anti-leishmania antibodies (IgG1, IgG2a, IgG2b and total IgG) were detected by ELISA assay. .. For isotype determination, goat anti-mouse IgG1, IgG2a and IgG2b (Sigma, USA) and HRP-conjugated mouse anti-goat antibody were used according to the manufacture’s instruction.

    Article Title: Expression and Characterization of Yeast Derived Chikungunya Virus Like Particles (CHIK-VLPs) and Its Evaluation as a Potential Vaccine Candidate
    Article Snippet: Briefly, CHIKV-VLP or inactivated CHIKV coated ELISA plate was incubated with 100μl (1:1000) of immunized sera and incubated for 1 h at 37°C followed by three washing with PBST. .. Wells were incubated with 100μl (1:1000) of goat anti-mouse IgG specific for each subtype (IgG1, IgG2a, IgG2b, IgG3) (Sigma, USA), at 37°C for 1 h. Following washing, 1:5000 dilution of rabbit anti-goat IgG HRP conjugate (Sigma, USA) was added and incubated at 37°C for 30 min.

    Article Title: Deletion of Fc? Receptor IIB Renders H-2b Mice Susceptible to Collagen-induced Arthritis
    Article Snippet: Serum antibody titers were measured by modification of an ELISA assay described previously ( ). .. The wells were washed three times with PBS containing 0.05% Tween 20, incubated with 50 μl of a 1:200 dilution of goat anti–mouse IgG1, IgG2a, IgG2b, or IgM coupled to horseradish peroxidase ( Sigma Chemical Co. ) at 4°C for 2 h, washed three times with PBS containing 0.05% Tween 20, and developed at room temperature for 30 min with 0.1 ml of TrueBlue Peroxidase Substrate (Kirkegaard & Perry Labs).

    Article Title: An efficient system to generate monoclonal antibodies against membrane-associated proteins by immunisation with antigen-expressing mammalian cells
    Article Snippet: .. IgG ELISA screen Maxisorp™plates (Nunc) were coated overnight at 4°C in a humid box with 100 μl of 5 μg/ml goat anti-mouse IgG (γ-chain specific) mAb (Sigma) diluted in PBS. .. After two washings with PBS containing 0.05% Tween-20, wells were blocked with blocking buffer (50 mM Tris, 140 mM NaCl, 5 mM EDTA, 0.05% NONidet P40, 0.25% gelatine, 1% BSA) for 1 h at 37°C and afterwards washed two times.

    Article Title: Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses
    Article Snippet: In brief, ELISA plates (Greiner, Germany) were coated with 280 ng purified F1S1 and blocked by 1% BSA in PBS (BSA-PBS). .. Goat anti-mouse IgG, rabbit anti-mouse IgG1, rabbit anti-mouse IgG2a, or goat anti-mouse IgA antibodies (Sigma, Germany), all conjugated to horseradish peroxidase, were used as the secondary antibodies.

    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
    Article Snippet: Paragraph title: ELISA. ... In order to evaluate the IgG subclass responses to PfCelTOS, the test was performed as described above, and instead of goat anti-mouse IgG-HRP, 100 μl of either goat anti-mouse IgG1, IgG2a, IgG2b, or IgG3 (diluted 1:1,000 in PBS-T buffer) (Sigma-Aldrich) as a secondary antibody was added to each well and incubated at RT for 1 h. After washing, the plates were incubated with a 1:10,000 dilution of anti-goat IgG-HRP (Sigma-Aldrich) at RT for 1 h, and the reaction was then developed by using an enzyme-specific substrate, as mentioned above.

    Article Title: G2 Dendrimer as a Carrier Can Enhance Immune Responses Against HCV-NS3 Protein in BALB/c Mice
    Article Snippet: Briefly, purified rNS3 (3 μg/ml ) was used as a captured molecule to coat 96-well polyvinyl chloride ELISA plates (Nunc, Denmark) for overnight at 4°C . .. IgG antibody subclasses were analyzed as mentioned above using goat anti-mouse IgG1, IgG2a, IgG2b antibodies (Sigma, Aldrich) and rabbit anti-goat IgG-HRP conjugate (Sigma, Aldrich).

    Incubation:

    Article Title: Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice
    Article Snippet: .. After three washes with PBS, the strips were further incubated for 2 h with goat anti-mouse IgG (Sigma-Aldrich) diluted 1:500 or goat anti-feline IgG (Southern Biotech) diluted 1:1000. ..

    Article Title: Interleukin-22 as a molecular adjuvant facilitates IL-17-producing CD8+ T cell responses against a HBV DNA vaccine in mice
    Article Snippet: .. After 5 time washes with PBST, 1:1000 diluted goat anti-mouse IgG, IgG1, or IgG2a conjugated with HPR was added to the wells, and the plates were incubated at 37 °C for 1 h. After five times of wash, 100 μl TMB buffer (Sigma, cat.no.T2885, 10 mg TMB powder dissolved in 200 μl H2 O and 800 μl ethanol) was added to each well. ..

    Article Title: Mechanistic Insight into the TH1-Biased Immune Response to Recombinant Subunit Vaccines Delivered by Probiotic Bacteria-Derived Outer Membrane Vesicles
    Article Snippet: ELISA for Serum Antibody Response Polystyrene microtiter 96-well plates (Maxisorp; Nunc Nalgene) were coated with GFP (5 µg/ml in carbonate buffer, pH 9.6) and incubated overnight at 4°C. .. Plates were washed six times with PBST, and biotinilated goat anti-mouse IgG, IgM (Sigma), or monoclonal IgG1 or IgG2 (BD Pharmingen) were added to the wells (1 µg/ml) for 1 hour at 37°C.

    Article Title: Heat Shock Proteins Enriched-Promastigotes of Leishmania major Inducing Th2 Immune Response in BALB/c Mice
    Article Snippet: Plates were blocked with 2% BSA in PBS at 37ºC for 1 h. Sera were diluted 1/200 and 100 µl was added to each well and incubated at 37°C for 30 min. After washing five times, 100 µl of HRP-conjugated rabbit anti-mouse (Sigma, USA) was added to each well and after 2 h, the plates were washed five times and 100 µl of TMB (3,3', 5,5"-tetra-methylbenzidine) substrate (KEM-EN-TEC, Denmark) was added to each well and incubated in the dark at 37C for 30 min. .. For isotype determination, goat anti-mouse IgG1, IgG2a and IgG2b (Sigma, USA) and HRP-conjugated mouse anti-goat antibody were used according to the manufacture’s instruction.

    Article Title: Expression and Characterization of Yeast Derived Chikungunya Virus Like Particles (CHIK-VLPs) and Its Evaluation as a Potential Vaccine Candidate
    Article Snippet: .. Wells were incubated with 100μl (1:1000) of goat anti-mouse IgG specific for each subtype (IgG1, IgG2a, IgG2b, IgG3) (Sigma, USA), at 37°C for 1 h. Following washing, 1:5000 dilution of rabbit anti-goat IgG HRP conjugate (Sigma, USA) was added and incubated at 37°C for 30 min. ..

    Article Title: Deletion of Fc? Receptor IIB Renders H-2b Mice Susceptible to Collagen-induced Arthritis
    Article Snippet: .. The wells were washed three times with PBS containing 0.05% Tween 20, incubated with 50 μl of a 1:200 dilution of goat anti–mouse IgG1, IgG2a, IgG2b, or IgM coupled to horseradish peroxidase ( Sigma Chemical Co. ) at 4°C for 2 h, washed three times with PBS containing 0.05% Tween 20, and developed at room temperature for 30 min with 0.1 ml of TrueBlue Peroxidase Substrate (Kirkegaard & Perry Labs). ..

    Article Title: Intradermal delivery of a fractional dose of influenza H7N9 split vaccine elicits protective immunity in mice and rats
    Article Snippet: Briefly, 4 HA units were mixed with serum for 30 min at room temperature in 96-well plates, 0.5% (v/v) turkey erythrocytes were added, and the samples were incubated for 40 min. .. Bound antibodies were detected using 1:20,000 dilutions of goat anti-mouse IgG, IgG1, and IgG2a (Sigma, USA) and goat-anti-rat IgG (Sigma), conjugated to horseradish peroxidase.

    Article Title: An efficient system to generate monoclonal antibodies against membrane-associated proteins by immunisation with antigen-expressing mammalian cells
    Article Snippet: IgG ELISA screen Maxisorp™plates (Nunc) were coated overnight at 4°C in a humid box with 100 μl of 5 μg/ml goat anti-mouse IgG (γ-chain specific) mAb (Sigma) diluted in PBS. .. 50 μl hybridoma supernatants were added to the wells and incubated for 1 h at 37°C.

    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
    Article Snippet: .. In order to evaluate the IgG subclass responses to PfCelTOS, the test was performed as described above, and instead of goat anti-mouse IgG-HRP, 100 μl of either goat anti-mouse IgG1, IgG2a, IgG2b, or IgG3 (diluted 1:1,000 in PBS-T buffer) (Sigma-Aldrich) as a secondary antibody was added to each well and incubated at RT for 1 h. After washing, the plates were incubated with a 1:10,000 dilution of anti-goat IgG-HRP (Sigma-Aldrich) at RT for 1 h, and the reaction was then developed by using an enzyme-specific substrate, as mentioned above. .. Antibody titration was done to assess the endpoint titers of anti-PfCelTOS raised in mice on days 38 and 180 after the first immunization by an ELISA.

    Article Title: G2 Dendrimer as a Carrier Can Enhance Immune Responses Against HCV-NS3 Protein in BALB/c Mice
    Article Snippet: After washing and blocking steps, wells were probed with optimum dilution of mice sera (1/1000 for total IgG), incubated for 1 hr , washed and further incubated with HRP-conjugated goat anti-mouse IgG (Sigma, Aldrich) as secondary antibody. .. IgG antibody subclasses were analyzed as mentioned above using goat anti-mouse IgG1, IgG2a, IgG2b antibodies (Sigma, Aldrich) and rabbit anti-goat IgG-HRP conjugate (Sigma, Aldrich).

    Article Title: B cells establish, but do not maintain, long-lived murine anti-peanut IgE
    Article Snippet: Ninety-six-well PVDF membrane ELISPOT plates (Millipore) were coated with CPE, goat anti-mouse IgG (Sigma), or rat anti-mouse IgE (R35-72, BD Biosciences). .. To enumerate 1) IgG antibody secreting cells (ASCs), 0.5×105 cells per well were incubated for 4 hours at 37°C; and 2) IgE ASCs, 1×106 cells per well were incubated overnight for 18 hours at 37°C.

    Enzyme-linked Immunospot:

    Article Title: B cells establish, but do not maintain, long-lived murine anti-peanut IgE
    Article Snippet: .. Ninety-six-well PVDF membrane ELISPOT plates (Millipore) were coated with CPE, goat anti-mouse IgG (Sigma), or rat anti-mouse IgE (R35-72, BD Biosciences). .. To enumerate 1) IgG antibody secreting cells (ASCs), 0.5×105 cells per well were incubated for 4 hours at 37°C; and 2) IgE ASCs, 1×106 cells per well were incubated overnight for 18 hours at 37°C.

    Stripping Membranes:

    Article Title: Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice
    Article Snippet: One strip containing S. schenckii ATCC 16345 CWPs was incubated with anti-rSsEno. .. After three washes with PBS, the strips were further incubated for 2 h with goat anti-mouse IgG (Sigma-Aldrich) diluted 1:500 or goat anti-feline IgG (Southern Biotech) diluted 1:1000.

    Infection:

    Article Title: Heat Shock Proteins Enriched-Promastigotes of Leishmania major Inducing Th2 Immune Response in BALB/c Mice
    Article Snippet: Ten days after final immunization and two months after infection, mice sera from each group were collected and anti-leishmania antibodies (IgG1, IgG2a, IgG2b and total IgG) were detected by ELISA assay. .. For isotype determination, goat anti-mouse IgG1, IgG2a and IgG2b (Sigma, USA) and HRP-conjugated mouse anti-goat antibody were used according to the manufacture’s instruction.

    Staining:

    Article Title: Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice
    Article Snippet: Two gels were stained with Coomassie brilliant blue R250, and the other gels were transferred to 0.45-μm-nitrocellulose membranes (GE Healthcare) using a mini Tank VEP-2 electroblotting system (Owl Separation Systems, Thermo Scientific) at 50 mM for 3 h. The membrane-cut strips were saturated with 5% dried skim milk in PBS for 4 h at 37 °C, and the strips containing rSsEno were incubated overnight at room temperature (RT) with anti-rSsEno serum (obtained from BALB/c mice seven days after being immunized subcutaneously twice at 14 day intervals with 100 µg rSsEno emulsified with Freund´s adjuvant) or sera from cats with confirmed sporotrichosis* (n = 34) obtained from the Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos (Lapclin-Dermzoo)/Instituto Nacional de Infectologia Evandro Chagas (INI)/Fundação Oswaldo Cruz, Rio de Janeiro, Brazil. .. After three washes with PBS, the strips were further incubated for 2 h with goat anti-mouse IgG (Sigma-Aldrich) diluted 1:500 or goat anti-feline IgG (Southern Biotech) diluted 1:1000.

    Article Title: Intradermal delivery of a fractional dose of influenza H7N9 split vaccine elicits protective immunity in mice and rats
    Article Snippet: Bound antibodies were detected using 1:20,000 dilutions of goat anti-mouse IgG, IgG1, and IgG2a (Sigma, USA) and goat-anti-rat IgG (Sigma), conjugated to horseradish peroxidase. .. Plates were stained with 3,3'5,5' tetramethyl benzidine substrate (Sigma), the reaction was stopped by adding 2 M H2 SO4 , and the optical density (OD) at 450 nm was read.

    Modification:

    Article Title: Deletion of Fc? Receptor IIB Renders H-2b Mice Susceptible to Collagen-induced Arthritis
    Article Snippet: Serum antibody titers were measured by modification of an ELISA assay described previously ( ). .. The wells were washed three times with PBS containing 0.05% Tween 20, incubated with 50 μl of a 1:200 dilution of goat anti–mouse IgG1, IgG2a, IgG2b, or IgM coupled to horseradish peroxidase ( Sigma Chemical Co. ) at 4°C for 2 h, washed three times with PBS containing 0.05% Tween 20, and developed at room temperature for 30 min with 0.1 ml of TrueBlue Peroxidase Substrate (Kirkegaard & Perry Labs).

    Western Blot:

    Article Title: Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice
    Article Snippet: Paragraph title: SDS-PAGE, western blot analysis ... After three washes with PBS, the strips were further incubated for 2 h with goat anti-mouse IgG (Sigma-Aldrich) diluted 1:500 or goat anti-feline IgG (Southern Biotech) diluted 1:1000.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Interleukin-22 as a molecular adjuvant facilitates IL-17-producing CD8+ T cell responses against a HBV DNA vaccine in mice
    Article Snippet: .. After 5 time washes with PBST, 1:1000 diluted goat anti-mouse IgG, IgG1, or IgG2a conjugated with HPR was added to the wells, and the plates were incubated at 37 °C for 1 h. After five times of wash, 100 μl TMB buffer (Sigma, cat.no.T2885, 10 mg TMB powder dissolved in 200 μl H2 O and 800 μl ethanol) was added to each well. ..

    Binding Assay:

    Article Title: B cells establish, but do not maintain, long-lived murine anti-peanut IgE
    Article Snippet: Ninety-six-well PVDF membrane ELISPOT plates (Millipore) were coated with CPE, goat anti-mouse IgG (Sigma), or rat anti-mouse IgE (R35-72, BD Biosciences). .. The following Ab were incubated sequentially for one hour at 37°C: biotin goat-anti-mouse IgG (BioLegend) and avidin-peroxidase for IgG ASCs; or sheep anti-mouse IgE (The Binding Site Group, Ltd), rabbit anti-sheep IgG and peroxidase goat anti-rabbit IgG (both from Jackson ImmunoResearch Laboratories Inc.) for IgE ASCs.

    Immunofluorescence:

    Article Title: A Novel Cellular Protein, p60, Interacting with both Herpes Simplex Virus 1 Regulatory Proteins ICP22 and ICP0 Is Modified in a Cell-Type-Specific Manner and Is Recruited to the Nucleus after Infection
    Article Snippet: Paragraph title: Immunofluorescence. ... The cells were rinsed three times with PBS and then reacted for 1 h at room temperature with goat anti-rabbit immunoglobulin G (IgG) conjugated to Texas red (Molecular Probes) and goat anti-mouse IgG conjugated to fluorescein isothiocyanate (FITC; Sigma).

    Blocking Assay:

    Article Title: An efficient system to generate monoclonal antibodies against membrane-associated proteins by immunisation with antigen-expressing mammalian cells
    Article Snippet: IgG ELISA screen Maxisorp™plates (Nunc) were coated overnight at 4°C in a humid box with 100 μl of 5 μg/ml goat anti-mouse IgG (γ-chain specific) mAb (Sigma) diluted in PBS. .. After two washings with PBS containing 0.05% Tween-20, wells were blocked with blocking buffer (50 mM Tris, 140 mM NaCl, 5 mM EDTA, 0.05% NONidet P40, 0.25% gelatine, 1% BSA) for 1 h at 37°C and afterwards washed two times.

    Article Title: G2 Dendrimer as a Carrier Can Enhance Immune Responses Against HCV-NS3 Protein in BALB/c Mice
    Article Snippet: After washing and blocking steps, wells were probed with optimum dilution of mice sera (1/1000 for total IgG), incubated for 1 hr , washed and further incubated with HRP-conjugated goat anti-mouse IgG (Sigma, Aldrich) as secondary antibody. .. IgG antibody subclasses were analyzed as mentioned above using goat anti-mouse IgG1, IgG2a, IgG2b antibodies (Sigma, Aldrich) and rabbit anti-goat IgG-HRP conjugate (Sigma, Aldrich).

    Software:

    Article Title: Heat Shock Proteins Enriched-Promastigotes of Leishmania major Inducing Th2 Immune Response in BALB/c Mice
    Article Snippet: For isotype determination, goat anti-mouse IgG1, IgG2a and IgG2b (Sigma, USA) and HRP-conjugated mouse anti-goat antibody were used according to the manufacture’s instruction. .. The data were analyzed with SPSS V.13 software by one-way ANOVA followed by Tukey's test.

    Article Title: Subcutaneous administration of a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis induces mucosal and systemic immune responses
    Article Snippet: Goat anti-mouse IgG, rabbit anti-mouse IgG1, rabbit anti-mouse IgG2a, or goat anti-mouse IgA antibodies (Sigma, Germany), all conjugated to horseradish peroxidase, were used as the secondary antibodies. .. Goat anti-mouse IgG, rabbit anti-mouse IgG1, rabbit anti-mouse IgG2a, or goat anti-mouse IgA antibodies (Sigma, Germany), all conjugated to horseradish peroxidase, were used as the secondary antibodies.

    Article Title: A Novel Cellular Protein, p60, Interacting with both Herpes Simplex Virus 1 Regulatory Proteins ICP22 and ICP0 Is Modified in a Cell-Type-Specific Manner and Is Recruited to the Nucleus after Infection
    Article Snippet: The cells were rinsed three times with PBS and then reacted for 1 h at room temperature with goat anti-rabbit immunoglobulin G (IgG) conjugated to Texas red (Molecular Probes) and goat anti-mouse IgG conjugated to fluorescein isothiocyanate (FITC; Sigma). .. The slides were examined with a Zeiss confocal fluorescence microscope, and digitized images were acquired with software provided by the manufacturer and printed with a Tektronix 440 phaser printer.

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  • 88
    Millipore rabbit monoclonal anti inos
    <t>NPY</t> inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on <t>iNOS</t> (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated
    Rabbit Monoclonal Anti Inos, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti inos/product/Millipore
    Average 88 stars, based on 1 article reviews
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    rabbit monoclonal anti inos - by Bioz Stars, 2020-02
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    86
    Millipore goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate
    Virion-specific <t>IgG</t> and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice
    Goat Anti Mouse Immunoglobulin G Igg Horseradish Peroxidase Conjugate, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate/product/Millipore
    Average 86 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate - by Bioz Stars, 2020-02
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    90
    Millipore mouse anti bax monoclonal
    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of <t>pro-caspase-8/9/3</t> (A) and <t>Bcl-2/Bax</t> (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P
    Mouse Anti Bax Monoclonal, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    NPY inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on iNOS (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated

    Journal: The Journal of Biological Chemistry

    Article Title: Neuropeptide Y Modulation of Interleukin-1? (IL-1?)-induced Nitric Oxide Production in Microglia *

    doi: 10.1074/jbc.M110.164020

    Figure Lengend Snippet: NPY inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on iNOS (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated

    Article Snippet: Antibodies used were as follows: rabbit polyclonal anti-NPY (1:1000) (Sigma); sheep polyclonal anti-Y1 R (1:1000) (AbD Serotec, Oxfordshire, UK); rabbit monoclonal anti-iNOS (1:250) (Millipore Corp., Bedford, MA); rat monoclonal anti-CD11b (1:1000) (AbD Serotec); rabbit monoclonal anti-NF-κB p65 (1:100) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) in 0.1% Triton X-100, 0.3% BSA solution; and Alexa Fluor 594 goat anti-rabbit, Alexa Fluor 594 donkey anti-sheep, Alexa Fluor 594 goat anti-rat, Alexa Fluor 488 donkey anti-rabbit, and Alexa Fluor 488 goat anti-rat (all 1:200 in PBS, from Molecular Probes, Eugene, OR).

    Techniques: Expressing

    NPY inhibits IL-1β-induced iNOS protein levels. A , confocal microscopy photomicrographs illustrate microglial cells treated with 1 μ m NPY and 1.5 ng/ml IL-1β for 6 h to assess the role of NPY and IL-1β in iNOS synthesis.

    Journal: The Journal of Biological Chemistry

    Article Title: Neuropeptide Y Modulation of Interleukin-1? (IL-1?)-induced Nitric Oxide Production in Microglia *

    doi: 10.1074/jbc.M110.164020

    Figure Lengend Snippet: NPY inhibits IL-1β-induced iNOS protein levels. A , confocal microscopy photomicrographs illustrate microglial cells treated with 1 μ m NPY and 1.5 ng/ml IL-1β for 6 h to assess the role of NPY and IL-1β in iNOS synthesis.

    Article Snippet: Antibodies used were as follows: rabbit polyclonal anti-NPY (1:1000) (Sigma); sheep polyclonal anti-Y1 R (1:1000) (AbD Serotec, Oxfordshire, UK); rabbit monoclonal anti-iNOS (1:250) (Millipore Corp., Bedford, MA); rat monoclonal anti-CD11b (1:1000) (AbD Serotec); rabbit monoclonal anti-NF-κB p65 (1:100) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) in 0.1% Triton X-100, 0.3% BSA solution; and Alexa Fluor 594 goat anti-rabbit, Alexa Fluor 594 donkey anti-sheep, Alexa Fluor 594 goat anti-rat, Alexa Fluor 488 donkey anti-rabbit, and Alexa Fluor 488 goat anti-rat (all 1:200 in PBS, from Molecular Probes, Eugene, OR).

    Techniques: Confocal Microscopy

    Virion-specific IgG and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice

    Journal:

    Article Title: Systemic Priming-Boosting Immunization with a Trivalent Plasmid DNA and Inactivated Murine Cytomegalovirus (MCMV) Vaccine Provides Long-Term Protection against Viral Replication following Systemic or Mucosal MCMV Challenge

    doi: 10.1128/JVI.79.1.159-175.2005

    Figure Lengend Snippet: Virion-specific IgG and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice

    Article Snippet: To detect gB, blocked blots were incubated with the monoclonal antibody 2E8.12A (a gift from Lambert Loh, University of Saskatchewan, Saskatoon, Canada), and bound antibody was detected using a goat-anti-mouse immunoglobulin G (IgG) horseradish peroxidase conjugate (Calbiochem) and enhanced chemiluminescence (Supersignal West Pico; Pierce).

    Techniques: Mouse Assay

    Virion-specific IgG and neutralizing-antibody responses in vaccinated mice. On the weeks of the experiment shown in Fig. , four to eight mice per vaccine group were retroorbitally bled and sera were prepared. Arrows and numbers indicate

    Journal:

    Article Title: Systemic Priming-Boosting Immunization with a Trivalent Plasmid DNA and Inactivated Murine Cytomegalovirus (MCMV) Vaccine Provides Long-Term Protection against Viral Replication following Systemic or Mucosal MCMV Challenge

    doi: 10.1128/JVI.79.1.159-175.2005

    Figure Lengend Snippet: Virion-specific IgG and neutralizing-antibody responses in vaccinated mice. On the weeks of the experiment shown in Fig. , four to eight mice per vaccine group were retroorbitally bled and sera were prepared. Arrows and numbers indicate

    Article Snippet: To detect gB, blocked blots were incubated with the monoclonal antibody 2E8.12A (a gift from Lambert Loh, University of Saskatchewan, Saskatoon, Canada), and bound antibody was detected using a goat-anti-mouse immunoglobulin G (IgG) horseradish peroxidase conjugate (Calbiochem) and enhanced chemiluminescence (Supersignal West Pico; Pierce).

    Techniques: Mouse Assay

    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Cell Culture, Transfection, Western Blot, Positive Control

    Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Activation Assay, Expressing, Cell Culture, Transfection, Western Blot

    Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Western Blot