alkaline phosphatase conjugated goat anti mouse igg  (SouthernBiotech)

 
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    Name:
    Goat F ab 2 Anti Mouse Kappa BIOT
    Description:

    Catalog Number:
    1052-08
    Price:
    None
    Source:
    Pepsin digest of Goat Anti-Mouse Kappa (SB Cat. No. 1050)
    Applications:
    Quality tested applications for relevant formats include -ELISA 1Flow CytometryOther referenced applications for relevant formats include -Immunohistochemistry-Paraffin Sections 8ELISpot 1 Stimulation 2-7
    Format:
    BIOT (Biotin)
    Isotype:
    Goat F(ab')2 IgG
    Buy from Supplier


    Structured Review

    SouthernBiotech alkaline phosphatase conjugated goat anti mouse igg
    Goat F ab 2 Anti Mouse Kappa BIOT

    https://www.bioz.com/result/alkaline phosphatase conjugated goat anti mouse igg/product/SouthernBiotech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alkaline phosphatase conjugated goat anti mouse igg - by Bioz Stars, 2021-06
    86/100 stars

    Images

    1) Product Images from "CD103+CD11b+ mucosal classical dendritic cells initiate long-term switched antibody responses to flagellin"

    Article Title: CD103+CD11b+ mucosal classical dendritic cells initiate long-term switched antibody responses to flagellin

    Journal: Mucosal immunology

    doi: 10.1038/mi.2017.105

    DP cDC control the induction of the mucosal Ab response to sFliC. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized (N.I) or primed and boosted with sFliC. The Ab response was evaluated 4 d post-boost. (a) Representative plots and absolute number (graphs) of plasma cells (TCRb - CD19 + B220 low CD138 + ) in the MLN. (b) ELISPOT analysis of sFliC IgG and IgA responses in the MLN and SI-LP. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are mean +SD (n=4 mice/group) and are from 1 representative experiment of three performed. *** p
    Figure Legend Snippet: DP cDC control the induction of the mucosal Ab response to sFliC. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized (N.I) or primed and boosted with sFliC. The Ab response was evaluated 4 d post-boost. (a) Representative plots and absolute number (graphs) of plasma cells (TCRb - CD19 + B220 low CD138 + ) in the MLN. (b) ELISPOT analysis of sFliC IgG and IgA responses in the MLN and SI-LP. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are mean +SD (n=4 mice/group) and are from 1 representative experiment of three performed. *** p

    Techniques Used: Mouse Assay, Enzyme-linked Immunospot

    The mucosal response to sFliC contributes to the systemic antibody response. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized or sFliC prime-boosted. (a) ELISPOT analysis of sFliC IgG and IgA responses in the BM. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are shown as mean +SD (n=12 mice/group) of three independent experiments pooled together. *** p
    Figure Legend Snippet: The mucosal response to sFliC contributes to the systemic antibody response. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized or sFliC prime-boosted. (a) ELISPOT analysis of sFliC IgG and IgA responses in the BM. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are shown as mean +SD (n=12 mice/group) of three independent experiments pooled together. *** p

    Techniques Used: Mouse Assay, Enzyme-linked Immunospot

    2) Product Images from "In Vivo Gene Transfer Using a Nonprimate Lentiviral Vector Pseudotyped with Ross River Virus Glycoproteins"

    Article Title: In Vivo Gene Transfer Using a Nonprimate Lentiviral Vector Pseudotyped with Ross River Virus Glycoproteins

    Journal: Journal of Virology

    doi: 10.1128/JVI.76.18.9378-9388.2002

    Antibody immune responses against β-galactosidase and the viral protein (p24) were comparable in mice receiving RRV/FIV vector or VSV-G/FIV vector and unrelated to the extent of Kupffer cell transduction. (A) Summary of the relationship between the percentages of transduced Kupffer cells and the production of antibody immune responses against β-galactosidase. The y axis represents the percentages of transduced Kupffer cells. The left-hand x axis represents mice that did not have antibody immune responses against β-galactosidase. The right-hand x axis indicates mice that developed antibody immune responses against β-galactosidase. Each symbol represents an individual animal. (B) Mice were intravenously injected via the tail vein with a single bolus of control buffer, 10 × 10 7 TU of RRV/FIV vector, or 35 × 10 7 TU of VSV-G/FIV vector. Serum samples were collected 3 weeks postinjection. β-Galactosidase-specific (left panel) and p24-specific (right panel) IgG and IgM antibody production were measured as described in Materials and Methods. Note that the scale of serum dilutions for β-galactosidase-specific antibodies is 2 logs higher than that for p24 specific antibodies.
    Figure Legend Snippet: Antibody immune responses against β-galactosidase and the viral protein (p24) were comparable in mice receiving RRV/FIV vector or VSV-G/FIV vector and unrelated to the extent of Kupffer cell transduction. (A) Summary of the relationship between the percentages of transduced Kupffer cells and the production of antibody immune responses against β-galactosidase. The y axis represents the percentages of transduced Kupffer cells. The left-hand x axis represents mice that did not have antibody immune responses against β-galactosidase. The right-hand x axis indicates mice that developed antibody immune responses against β-galactosidase. Each symbol represents an individual animal. (B) Mice were intravenously injected via the tail vein with a single bolus of control buffer, 10 × 10 7 TU of RRV/FIV vector, or 35 × 10 7 TU of VSV-G/FIV vector. Serum samples were collected 3 weeks postinjection. β-Galactosidase-specific (left panel) and p24-specific (right panel) IgG and IgM antibody production were measured as described in Materials and Methods. Note that the scale of serum dilutions for β-galactosidase-specific antibodies is 2 logs higher than that for p24 specific antibodies.

    Techniques Used: Mouse Assay, Plasmid Preparation, Transduction, Injection

    3) Product Images from "Anti-Islet Autoantibodies Trigger Autoimmune Diabetes in the Presence of an Increased Frequency of Islet-Reactive CD4 T Cells"

    Article Title: Anti-Islet Autoantibodies Trigger Autoimmune Diabetes in the Presence of an Increased Frequency of Islet-Reactive CD4 T Cells

    Journal: Diabetes

    doi: 10.2337/db10-1344

    Anti-HEL autoantibodies enhance accumulation of HEL-specific CD4+ T cells. Splenocytes from TCR+ mice (bearing the CD45.1 allelic marker) were labeled with the cell division dye CFSE and injected intravenously into 24 HEL+ recipients (CD45.2) on day 0. Twelve recipients were given 200 μL HEL-immune serum and twelve were given 200 μL control serum i.p. starting on day −1 and on alternate days thereafter in surviving mice. Flow cytometry on recipient spleens was performed on days 3, 5, 7, and 10, each time using three mice from each treatment group plus two control nontransgenic mice that had received cells but no serum injections. A : Representative flow cytometric plots (from day 5 after transfer) showing gates used to track donor-derived CD4 + cells and B220 + “tracer” B cells ( upper panel ), plus TCR HEL expression and CFSE dilution on the donor-derived CD4 + cells ( lower panel ). B : Graphs show the ratio of donor CD4 + cells to donor B220 + cells, and the shaded areas represent the proportion of TCR HEL + cells among donor-derived CD4 + cells. C : Percentage of CFSE-diluted cells among donor-derived CD4 + cells. In ( B ) and ( C ), symbols indicate group means; error bars show the SD. Statistical differences between HEL-immune and control serum groups were analyzed using a ratio t test (see research design and methods ). The data shown are from one of three experiments that yielded comparable results. D : The experiment described in A – C was repeated, except that the TCR+ donors were CD45.2+ Bim-deficient mice and the recipients were CD45.1+. Histogram ( top ) displays CFSE on donor (CD45.2+) CD4+ cells in HEL+ hosts that received HEL-immune or control serum, or in nontransgenic hosts as indicated, with a summary of the data ( bottom ) for multiple recipients. E : Diabetes incidence in TCR+HEL+ neonates injected on days 2, 4, and 6 after birth with serum from HEL-immunized mice and on days 1, 3, 5, 8, 11, 14, and 17 after birth with Fcγ R -blocking antibody or rat IgG2b isotype control. Statistical analysis used the log-rank method.
    Figure Legend Snippet: Anti-HEL autoantibodies enhance accumulation of HEL-specific CD4+ T cells. Splenocytes from TCR+ mice (bearing the CD45.1 allelic marker) were labeled with the cell division dye CFSE and injected intravenously into 24 HEL+ recipients (CD45.2) on day 0. Twelve recipients were given 200 μL HEL-immune serum and twelve were given 200 μL control serum i.p. starting on day −1 and on alternate days thereafter in surviving mice. Flow cytometry on recipient spleens was performed on days 3, 5, 7, and 10, each time using three mice from each treatment group plus two control nontransgenic mice that had received cells but no serum injections. A : Representative flow cytometric plots (from day 5 after transfer) showing gates used to track donor-derived CD4 + cells and B220 + “tracer” B cells ( upper panel ), plus TCR HEL expression and CFSE dilution on the donor-derived CD4 + cells ( lower panel ). B : Graphs show the ratio of donor CD4 + cells to donor B220 + cells, and the shaded areas represent the proportion of TCR HEL + cells among donor-derived CD4 + cells. C : Percentage of CFSE-diluted cells among donor-derived CD4 + cells. In ( B ) and ( C ), symbols indicate group means; error bars show the SD. Statistical differences between HEL-immune and control serum groups were analyzed using a ratio t test (see research design and methods ). The data shown are from one of three experiments that yielded comparable results. D : The experiment described in A – C was repeated, except that the TCR+ donors were CD45.2+ Bim-deficient mice and the recipients were CD45.1+. Histogram ( top ) displays CFSE on donor (CD45.2+) CD4+ cells in HEL+ hosts that received HEL-immune or control serum, or in nontransgenic hosts as indicated, with a summary of the data ( bottom ) for multiple recipients. E : Diabetes incidence in TCR+HEL+ neonates injected on days 2, 4, and 6 after birth with serum from HEL-immunized mice and on days 1, 3, 5, 8, 11, 14, and 17 after birth with Fcγ R -blocking antibody or rat IgG2b isotype control. Statistical analysis used the log-rank method.

    Techniques Used: Mouse Assay, Marker, Labeling, Injection, Flow Cytometry, Cytometry, Derivative Assay, Expressing, Blocking Assay

    Effect of passive transfer of antibody on diabetes in TCR+ HEL+ mice. A : Breeding strategy for maternal antibody transfer experiments. Nontransgenic B10.BR females were immunized with HEL or OVA protein emulsified in CFA to induce specific antibodies and then mated with TCR+HEL+ males to yield 25% TCR+HEL+ offspring and equal numbers of singly transgenic or nontransgenic littermates. B : Anti-HEL IgG titers in newborns from mothers immunized as illustrated in ( A ); 70-day-old pups from immunized or unimmunized mothers and mothers 5 weeks after immunization. C : Diabetes incidence in offspring with the indicated genotypes bred from HEL-immunized mothers. D : Diabetes incidence in TCR+HEL+ offspring of mothers immunized with either HEL (closed symbols) or OVA (open symbols). E and F : Diabetes incidence in TCR+HEL+ neonates injected on days 1, 3, and 5 after birth with serum ( E ) or purified IgG ( F ) from HEL- or OVA-immunized mice.
    Figure Legend Snippet: Effect of passive transfer of antibody on diabetes in TCR+ HEL+ mice. A : Breeding strategy for maternal antibody transfer experiments. Nontransgenic B10.BR females were immunized with HEL or OVA protein emulsified in CFA to induce specific antibodies and then mated with TCR+HEL+ males to yield 25% TCR+HEL+ offspring and equal numbers of singly transgenic or nontransgenic littermates. B : Anti-HEL IgG titers in newborns from mothers immunized as illustrated in ( A ); 70-day-old pups from immunized or unimmunized mothers and mothers 5 weeks after immunization. C : Diabetes incidence in offspring with the indicated genotypes bred from HEL-immunized mothers. D : Diabetes incidence in TCR+HEL+ offspring of mothers immunized with either HEL (closed symbols) or OVA (open symbols). E and F : Diabetes incidence in TCR+HEL+ neonates injected on days 1, 3, and 5 after birth with serum ( E ) or purified IgG ( F ) from HEL- or OVA-immunized mice.

    Techniques Used: Mouse Assay, Transgenic Assay, Injection, Purification

    High autoantibodies and requirement for B cells for accelerated progression to diabetes in Roquin san/san TCR+HEL+ mice. A and B : Titers of anti-HEL IgG ( A ) and IgG2a ( B ) in TCR+HEL+ mice of the indicated Roquin genotypes and diabetes status. C : Anti-HEL IgG titers in diabetic TCR+HEL+ mice of the indicated genotypes. D : Diabetes incidence in Roquin san/san TCR+HEL+ mice that were either B cell deficient ( Cd79a ken/ken ; filled circles) or B cell sufficient ( Cd79a +/+ ; triangles) and in Roquin +/+ Cd79a ken/ken TCR+HEL+ mice (open circles). E : Diabetes incidence in Roquin san/san TCR+HEL+ mice with or without an IgMD HEL transgene that prevents isotype switching of HEL-specific B cells, and control TCR+HEL+ mice with normal Roquin and B-cell repertoire. F : Diabetes incidence in Roquin san/+ TCR+HEL+ mice after transfer of a single 200 μL dose of serum from diabetic Roquin san/san TCR+HEL+ mice (closed symbols) or from diabetic Roquin san/+ or Roquin +/+ TCR+HEL+ mice (open symbols).
    Figure Legend Snippet: High autoantibodies and requirement for B cells for accelerated progression to diabetes in Roquin san/san TCR+HEL+ mice. A and B : Titers of anti-HEL IgG ( A ) and IgG2a ( B ) in TCR+HEL+ mice of the indicated Roquin genotypes and diabetes status. C : Anti-HEL IgG titers in diabetic TCR+HEL+ mice of the indicated genotypes. D : Diabetes incidence in Roquin san/san TCR+HEL+ mice that were either B cell deficient ( Cd79a ken/ken ; filled circles) or B cell sufficient ( Cd79a +/+ ; triangles) and in Roquin +/+ Cd79a ken/ken TCR+HEL+ mice (open circles). E : Diabetes incidence in Roquin san/san TCR+HEL+ mice with or without an IgMD HEL transgene that prevents isotype switching of HEL-specific B cells, and control TCR+HEL+ mice with normal Roquin and B-cell repertoire. F : Diabetes incidence in Roquin san/+ TCR+HEL+ mice after transfer of a single 200 μL dose of serum from diabetic Roquin san/san TCR+HEL+ mice (closed symbols) or from diabetic Roquin san/+ or Roquin +/+ TCR+HEL+ mice (open symbols).

    Techniques Used: Mouse Assay

    A maternally transmitted epigenetic factor increases diabetes incidence in the TCR+HEL+ offspring of formerly diabetic mothers. A : Diabetes incidence in TCR+HEL+ mice bred from TCR+HEL+ mothers or fathers that had spontaneously developed diabetes and had euglycemia stably restored by transplanting nontransgenic islets. The diabetes-cured TCR+HEL+ parents were bred with nontransgenic partners. Statistical analysis used the log-rank method. B : Diabetes incidence for the cohort described in ( A ) stratified by sex of the offspring. C : For each of 20 diabetes-cured TCR+HEL+ female breeders used to generate the cohort described in ( A ), two serial serum anti-HEL IgG titers (connected by a line) are plotted against the time of blood sampling relative to islet transplantation on day 0 ( left ). Serum anti-HEL IgG titers for 20 nondiabetic TCR+HEL+ females from the same colony as the diabetes-cured females ( right ).
    Figure Legend Snippet: A maternally transmitted epigenetic factor increases diabetes incidence in the TCR+HEL+ offspring of formerly diabetic mothers. A : Diabetes incidence in TCR+HEL+ mice bred from TCR+HEL+ mothers or fathers that had spontaneously developed diabetes and had euglycemia stably restored by transplanting nontransgenic islets. The diabetes-cured TCR+HEL+ parents were bred with nontransgenic partners. Statistical analysis used the log-rank method. B : Diabetes incidence for the cohort described in ( A ) stratified by sex of the offspring. C : For each of 20 diabetes-cured TCR+HEL+ female breeders used to generate the cohort described in ( A ), two serial serum anti-HEL IgG titers (connected by a line) are plotted against the time of blood sampling relative to islet transplantation on day 0 ( left ). Serum anti-HEL IgG titers for 20 nondiabetic TCR+HEL+ females from the same colony as the diabetes-cured females ( right ).

    Techniques Used: Mouse Assay, Stable Transfection, Sampling, Transplantation Assay

    4) Product Images from "Monoclonal antibodies detect receptor-induced binding sites in Glu-plasminogen"

    Article Title: Monoclonal antibodies detect receptor-induced binding sites in Glu-plasminogen

    Journal: Blood

    doi: 10.1182/blood-2010-11-316943

    Western blotting of anti-Pg mAbs. Glu-Pg, K1-3, K4, and K5-PD were each electrophoresed on 12.5% SDS gels under nonreducing conditions and Western blotted with the indicated mAbs (panels A-F). In controls, no bands were detected when normal mouse IgG
    Figure Legend Snippet: Western blotting of anti-Pg mAbs. Glu-Pg, K1-3, K4, and K5-PD were each electrophoresed on 12.5% SDS gels under nonreducing conditions and Western blotted with the indicated mAbs (panels A-F). In controls, no bands were detected when normal mouse IgG

    Techniques Used: Western Blot

    5) Product Images from "Immunization with an Apoptotic Cell-Binding Protein Recapitulates the Nephritis and Sequential Autoantibody Emergence of Systemic Lupus Erythematosus"

    Article Title: Immunization with an Apoptotic Cell-Binding Protein Recapitulates the Nephritis and Sequential Autoantibody Emergence of Systemic Lupus Erythematosus

    Journal:

    doi:

    LPS and TNF- α differentially affect the IgG and IgM anti- β 2 GPI responses to soluble and apoptotic cell-bound β 2 GPI. BALB/c mice were immunized with soluble or apoptotic cell-bound HEPES buffer without (−) or with (+) human
    Figure Legend Snippet: LPS and TNF- α differentially affect the IgG and IgM anti- β 2 GPI responses to soluble and apoptotic cell-bound β 2 GPI. BALB/c mice were immunized with soluble or apoptotic cell-bound HEPES buffer without (−) or with (+) human

    Techniques Used: Mouse Assay

    6) Product Images from "Optimized Mucosal Modified Vaccinia Virus Ankara Prime/Soluble gp120 Boost HIV Vaccination Regimen Induces Antibody Responses Similar to Those of an Intramuscular Regimen"

    Article Title: Optimized Mucosal Modified Vaccinia Virus Ankara Prime/Soluble gp120 Boost HIV Vaccination Regimen Induces Antibody Responses Similar to Those of an Intramuscular Regimen

    Journal: Journal of Virology

    doi: 10.1128/JVI.00475-19

    Intranasal prime with combined intranasal and intramuscular boosting induces serum antibodies and viral neutralization similar to those induced by the parenteral i.m. prime/i.m. boost regimen. Rhesus macaques ( n = 6/group) were vaccinated with the same i.n./i.m.-plus-i.n. or i.m./i.m. regimen described for rabbits. An additional booster vaccine was given in week 24. Serum was collected at the indicated time points, and antigen-specific antibody titers were determined by ELISA. Antibody responses were compared between the two vaccination regimens at each time point using a Mann-Whitney test. (A) Time course of serum gp120-specific IgG titers during the vaccination regimen. The i.m./i.m. group had significantly higher serum gp120-specific IgG titers after administration of the priming immunization (week 8), while the i.n./i.m.-plus-i.n. group had significantly higher titers 7 weeks after administration of the second booster vaccination (week 27) ( P = 0.01 and 0.03, respectively). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) Time course of serum gp120-specific IgA titers. The i.m./i.m. regimen resulted in significantly higher anti-gp120 IgA titers than the i.n./i.m.-plus-i.n. regimen at week 15 ( P = 0.04). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (C) Fecal gp120-specific IgG titers were not significantly different between vaccination regimens at any time point tested. The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (D) While all NHPs had undetectable fecal gp120-specific IgA at a 1:16 dilution at week 19, low levels of fecal gp120-specific IgA were detected in a few NHPs in each vaccination regimen on week 27. The difference between time points was not significant ( P = 0.6 for i.m./i.m.; P = 0.3 for i.n./i.m.-plus-i.n.). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (E) Neutralization of tier 1a (MW965.26) and tier 1b (all others) pseudoviruses was not significantly different between the vaccination regimens after both the 2nd and 3rd boosts. The horizontal lines represent the geometric mean titers.
    Figure Legend Snippet: Intranasal prime with combined intranasal and intramuscular boosting induces serum antibodies and viral neutralization similar to those induced by the parenteral i.m. prime/i.m. boost regimen. Rhesus macaques ( n = 6/group) were vaccinated with the same i.n./i.m.-plus-i.n. or i.m./i.m. regimen described for rabbits. An additional booster vaccine was given in week 24. Serum was collected at the indicated time points, and antigen-specific antibody titers were determined by ELISA. Antibody responses were compared between the two vaccination regimens at each time point using a Mann-Whitney test. (A) Time course of serum gp120-specific IgG titers during the vaccination regimen. The i.m./i.m. group had significantly higher serum gp120-specific IgG titers after administration of the priming immunization (week 8), while the i.n./i.m.-plus-i.n. group had significantly higher titers 7 weeks after administration of the second booster vaccination (week 27) ( P = 0.01 and 0.03, respectively). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) Time course of serum gp120-specific IgA titers. The i.m./i.m. regimen resulted in significantly higher anti-gp120 IgA titers than the i.n./i.m.-plus-i.n. regimen at week 15 ( P = 0.04). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (C) Fecal gp120-specific IgG titers were not significantly different between vaccination regimens at any time point tested. The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (D) While all NHPs had undetectable fecal gp120-specific IgA at a 1:16 dilution at week 19, low levels of fecal gp120-specific IgA were detected in a few NHPs in each vaccination regimen on week 27. The difference between time points was not significant ( P = 0.6 for i.m./i.m.; P = 0.3 for i.n./i.m.-plus-i.n.). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (E) Neutralization of tier 1a (MW965.26) and tier 1b (all others) pseudoviruses was not significantly different between the vaccination regimens after both the 2nd and 3rd boosts. The horizontal lines represent the geometric mean titers.

    Techniques Used: Neutralization, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Rabbit and NHP antibody responses to both prime/boost regimens were similar. New Zealand White rabbits and rhesus macaques received a priming immunization of MVAgp120 in week 0 and gp120 boosts in weeks 12 and 16. All the vaccines were either administered intramuscularly (i.m. prime/i.m. boost) or the MVAgp120 prime was administered intranasally and boosting consisted of both intramuscular and intranasal components (i.n./i.m.-plus-i.n.) ( n = 6/group). The animals were bled at the time points indicated, and antibody responses were determined by ELISA. (A) Serum gp120-specific IgG responses after administration of the MVAgp120 prime (weeks 6 to 8), the first gp120 boost (week 15), and the second gp120 boost (week 19). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) After administration of the second booster vaccine (week 19), rabbits and NHPs had similar serum V1V2-specific IgG and serum gp120-specific IgA responses. Fecal gp120-specific responses were also similar between the species. The horizontal lines represent the geometric mean titers.
    Figure Legend Snippet: Rabbit and NHP antibody responses to both prime/boost regimens were similar. New Zealand White rabbits and rhesus macaques received a priming immunization of MVAgp120 in week 0 and gp120 boosts in weeks 12 and 16. All the vaccines were either administered intramuscularly (i.m. prime/i.m. boost) or the MVAgp120 prime was administered intranasally and boosting consisted of both intramuscular and intranasal components (i.n./i.m.-plus-i.n.) ( n = 6/group). The animals were bled at the time points indicated, and antibody responses were determined by ELISA. (A) Serum gp120-specific IgG responses after administration of the MVAgp120 prime (weeks 6 to 8), the first gp120 boost (week 15), and the second gp120 boost (week 19). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) After administration of the second booster vaccine (week 19), rabbits and NHPs had similar serum V1V2-specific IgG and serum gp120-specific IgA responses. Fecal gp120-specific responses were also similar between the species. The horizontal lines represent the geometric mean titers.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Rabbits developed undetectable or low mucosal antibody responses to all of the prime/boost regimens. New Zealand White rabbits received a priming immunization of MVAgp120 either i.m. or i.n. in week 0 and gp120 boosts i.m. and/or i.n. in weeks 12 and 16. Feces were collected from all individuals ( n = 4 to 6/group), and vaginal lavage was performed on the females ( n = 3 or 4/group). The results shown are from week 19, 3 weeks after administration of the second booster vaccine. (A) Fifty to 100% of the rabbits in each group developed detectable fecal gp120-specific IgG titers. (B) Twenty-five to 50% of the rabbits in each group developed fecal gp120-specific IgA titers. (C) Vaginal gp120-specific IgG was detectable in 50 to 100% of the female rabbits in each group but tended to be lower than the fecal gp120-specific IgG titers observed. (D) Vaginal gp120-specific IgA was detectable in only 2 of the 4 i.m./i.n.-vaccinated females. The horizontal lines indicate the geometric mean titers.
    Figure Legend Snippet: Rabbits developed undetectable or low mucosal antibody responses to all of the prime/boost regimens. New Zealand White rabbits received a priming immunization of MVAgp120 either i.m. or i.n. in week 0 and gp120 boosts i.m. and/or i.n. in weeks 12 and 16. Feces were collected from all individuals ( n = 4 to 6/group), and vaginal lavage was performed on the females ( n = 3 or 4/group). The results shown are from week 19, 3 weeks after administration of the second booster vaccine. (A) Fifty to 100% of the rabbits in each group developed detectable fecal gp120-specific IgG titers. (B) Twenty-five to 50% of the rabbits in each group developed fecal gp120-specific IgA titers. (C) Vaginal gp120-specific IgG was detectable in 50 to 100% of the female rabbits in each group but tended to be lower than the fecal gp120-specific IgG titers observed. (D) Vaginal gp120-specific IgA was detectable in only 2 of the 4 i.m./i.n.-vaccinated females. The horizontal lines indicate the geometric mean titers.

    Techniques Used:

    7) Product Images from "Relationships among Dissemination of Primary Parainfluenza Virus Infection in the Respiratory Tract, Mucosal and Peripheral Immune Responses, and Protection from Reinfection: a Noninvasive Bioluminescence-Imaging Study"

    Article Title: Relationships among Dissemination of Primary Parainfluenza Virus Infection in the Respiratory Tract, Mucosal and Peripheral Immune Responses, and Protection from Reinfection: a Noninvasive Bioluminescence-Imaging Study

    Journal: Journal of Virology

    doi: 10.1128/JVI.03581-14

    Differential mucosal and systemic immunological responses to primary infection based on the inoculation method. Groups of mice were mock inoculated with PBS, intramuscularly inoculated with 1 × 10 6 PFU of rSeV-luc(M-F*) (IM), or intranasally (IN) inoculated with a low dose and a high volume (70 PFU in 5 μl) or a high dose and a high volume (7,000 PFU in 30 μl) of either rSeV-luc(M-F*) or rSeV-luc(P-M). (A and B) After 11 days of infection, mice were euthanized to determine the numbers of CD3 + T lymphocytes in whole lungs (A) or bronchoalveolar lavage fluid (B) by flow cytometry. Each data point for infiltrating T lymphocytes represents an individual mouse from triplicate experiments (PBS group, n = 8; i.m. group, n = 6; i.n. groups, n = 9), and horizontal bars show the group means. (C to F) For other groups, mice were euthanized after 28 days of infection to recover NALT and whole lungs to quantify IgA (C and D) and IgG (E and F) antibody-forming cells by an ELISPOT assay. Mucosal antibody-forming cell experiments were done in triplicate, and a representative data set is shown ( n = 3 with duplicate wells for each ELISPOT assay). (G and H) After 30 days of infection, peripheral blood was collected so that Sendai virus-specific binding (G) and neutralizing (H) antibody (Ab) titers in sera could be determined by reciprocal endpoint dilutions in ELISAs and microneutralization assays, respectively. Panel G displays individual values and means ± standard deviations; panel H displays means ± standard deviations [ n = 13 for the PBS group, n = 20 for the i.m. group, n = 24 for i.n. rSeV-luc(M-F*) at a high dose and volume, and n = 30 for the remaining three i.n. groups]. Significance was determined by using the Student t test. For panels A, D, and F, there was no statistically significant difference between the i.m. and low-dose and low-volume i.n. treatment groups. For panels B, C, E, G, and H, there is a statistically significant difference (*, P
    Figure Legend Snippet: Differential mucosal and systemic immunological responses to primary infection based on the inoculation method. Groups of mice were mock inoculated with PBS, intramuscularly inoculated with 1 × 10 6 PFU of rSeV-luc(M-F*) (IM), or intranasally (IN) inoculated with a low dose and a high volume (70 PFU in 5 μl) or a high dose and a high volume (7,000 PFU in 30 μl) of either rSeV-luc(M-F*) or rSeV-luc(P-M). (A and B) After 11 days of infection, mice were euthanized to determine the numbers of CD3 + T lymphocytes in whole lungs (A) or bronchoalveolar lavage fluid (B) by flow cytometry. Each data point for infiltrating T lymphocytes represents an individual mouse from triplicate experiments (PBS group, n = 8; i.m. group, n = 6; i.n. groups, n = 9), and horizontal bars show the group means. (C to F) For other groups, mice were euthanized after 28 days of infection to recover NALT and whole lungs to quantify IgA (C and D) and IgG (E and F) antibody-forming cells by an ELISPOT assay. Mucosal antibody-forming cell experiments were done in triplicate, and a representative data set is shown ( n = 3 with duplicate wells for each ELISPOT assay). (G and H) After 30 days of infection, peripheral blood was collected so that Sendai virus-specific binding (G) and neutralizing (H) antibody (Ab) titers in sera could be determined by reciprocal endpoint dilutions in ELISAs and microneutralization assays, respectively. Panel G displays individual values and means ± standard deviations; panel H displays means ± standard deviations [ n = 13 for the PBS group, n = 20 for the i.m. group, n = 24 for i.n. rSeV-luc(M-F*) at a high dose and volume, and n = 30 for the remaining three i.n. groups]. Significance was determined by using the Student t test. For panels A, D, and F, there was no statistically significant difference between the i.m. and low-dose and low-volume i.n. treatment groups. For panels B, C, E, G, and H, there is a statistically significant difference (*, P

    Techniques Used: Infection, Mouse Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunospot, Binding Assay

    8) Product Images from "Antibody and CD8+ T cell memory response to influenza A/PR/8/34 infection is reduced in treadmill-exercised mice, yet still protective"

    Article Title: Antibody and CD8+ T cell memory response to influenza A/PR/8/34 infection is reduced in treadmill-exercised mice, yet still protective

    Journal: Journal of Applied Physiology

    doi: 10.1152/japplphysiol.01355.2012

    Body weight loss, percent survival, and memory recall response to secondary infectious challenge with lethal-dose A/PR/8/34. Body weight loss ( A ) and percent survival ( B ) were determined for previously infected treadmill-exercised and nonexercised mice. Naïve mice included as a lethal dose (A/PR/8/34) control. Mice were monitored until day 7 post secondary challenge (PSC). C: serum anti-influenza IgG and subtypes (IgG1 and IgG2a) measured by indirect ELISA in the serum of all mice at day 8 PSC. ELISA results are expressed as mean optical density at 405 nm ± SE. Sera from noninfected mice were included as an ELISA control. Noninfected mice had significantly less IgG, IgG1, and IgG2a than No-Ex or TM-Ex mice ( P
    Figure Legend Snippet: Body weight loss, percent survival, and memory recall response to secondary infectious challenge with lethal-dose A/PR/8/34. Body weight loss ( A ) and percent survival ( B ) were determined for previously infected treadmill-exercised and nonexercised mice. Naïve mice included as a lethal dose (A/PR/8/34) control. Mice were monitored until day 7 post secondary challenge (PSC). C: serum anti-influenza IgG and subtypes (IgG1 and IgG2a) measured by indirect ELISA in the serum of all mice at day 8 PSC. ELISA results are expressed as mean optical density at 405 nm ± SE. Sera from noninfected mice were included as an ELISA control. Noninfected mice had significantly less IgG, IgG1, and IgG2a than No-Ex or TM-Ex mice ( P

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

    A and B: anti-influenza IgG and subtypes 14 and 28 days after challenge with an i.p. injection of live A/PR/8/34. IgG and subtypes detected in the serum at day 14 ( A ) and day 28 ( B ) post primary challenge by indirect ELISA. Noninfected mice had significantly less IgG, IgG1, and IgG2a than infected No-Ex or TM-EX mice at day 14 ( P
    Figure Legend Snippet: A and B: anti-influenza IgG and subtypes 14 and 28 days after challenge with an i.p. injection of live A/PR/8/34. IgG and subtypes detected in the serum at day 14 ( A ) and day 28 ( B ) post primary challenge by indirect ELISA. Noninfected mice had significantly less IgG, IgG1, and IgG2a than infected No-Ex or TM-EX mice at day 14 ( P

    Techniques Used: Injection, Indirect ELISA, Mouse Assay, Infection

    9) Product Images from "Optimized Mucosal Modified Vaccinia Virus Ankara Prime/Soluble gp120 Boost HIV Vaccination Regimen Induces Antibody Responses Similar to Those of an Intramuscular Regimen"

    Article Title: Optimized Mucosal Modified Vaccinia Virus Ankara Prime/Soluble gp120 Boost HIV Vaccination Regimen Induces Antibody Responses Similar to Those of an Intramuscular Regimen

    Journal: Journal of Virology

    doi: 10.1128/JVI.00475-19

    Intranasal prime with combined intranasal and intramuscular boosting induces serum antibodies and viral neutralization similar to those induced by the parenteral i.m. prime/i.m. boost regimen. Rhesus macaques ( n  = 6/group) were vaccinated with the same i.n./i.m.-plus-i.n. or i.m./i.m. regimen described for rabbits. An additional booster vaccine was given in week 24. Serum was collected at the indicated time points, and antigen-specific antibody titers were determined by ELISA. Antibody responses were compared between the two vaccination regimens at each time point using a Mann-Whitney test. (A) Time course of serum gp120-specific IgG titers during the vaccination regimen. The i.m./i.m. group had significantly higher serum gp120-specific IgG titers after administration of the priming immunization (week 8), while the i.n./i.m.-plus-i.n. group had significantly higher titers 7 weeks after administration of the second booster vaccination (week 27) ( P  = 0.01 and 0.03, respectively). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) Time course of serum gp120-specific IgA titers. The i.m./i.m. regimen resulted in significantly higher anti-gp120 IgA titers than the i.n./i.m.-plus-i.n. regimen at week 15 ( P  = 0.04). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (C) Fecal gp120-specific IgG titers were not significantly different between vaccination regimens at any time point tested. The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (D) While all NHPs had undetectable fecal gp120-specific IgA at a 1:16 dilution at week 19, low levels of fecal gp120-specific IgA were detected in a few NHPs in each vaccination regimen on week 27. The difference between time points was not significant ( P  = 0.6 for i.m./i.m.; P  = 0.3 for i.n./i.m.-plus-i.n.). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (E) Neutralization of tier 1a (MW965.26) and tier 1b (all others) pseudoviruses was not significantly different between the vaccination regimens after both the 2nd and 3rd boosts. The horizontal lines represent the geometric mean titers.
    Figure Legend Snippet: Intranasal prime with combined intranasal and intramuscular boosting induces serum antibodies and viral neutralization similar to those induced by the parenteral i.m. prime/i.m. boost regimen. Rhesus macaques ( n  = 6/group) were vaccinated with the same i.n./i.m.-plus-i.n. or i.m./i.m. regimen described for rabbits. An additional booster vaccine was given in week 24. Serum was collected at the indicated time points, and antigen-specific antibody titers were determined by ELISA. Antibody responses were compared between the two vaccination regimens at each time point using a Mann-Whitney test. (A) Time course of serum gp120-specific IgG titers during the vaccination regimen. The i.m./i.m. group had significantly higher serum gp120-specific IgG titers after administration of the priming immunization (week 8), while the i.n./i.m.-plus-i.n. group had significantly higher titers 7 weeks after administration of the second booster vaccination (week 27) ( P  = 0.01 and 0.03, respectively). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) Time course of serum gp120-specific IgA titers. The i.m./i.m. regimen resulted in significantly higher anti-gp120 IgA titers than the i.n./i.m.-plus-i.n. regimen at week 15 ( P  = 0.04). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (C) Fecal gp120-specific IgG titers were not significantly different between vaccination regimens at any time point tested. The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (D) While all NHPs had undetectable fecal gp120-specific IgA at a 1:16 dilution at week 19, low levels of fecal gp120-specific IgA were detected in a few NHPs in each vaccination regimen on week 27. The difference between time points was not significant ( P  = 0.6 for i.m./i.m.; P  = 0.3 for i.n./i.m.-plus-i.n.). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (E) Neutralization of tier 1a (MW965.26) and tier 1b (all others) pseudoviruses was not significantly different between the vaccination regimens after both the 2nd and 3rd boosts. The horizontal lines represent the geometric mean titers.

    Techniques Used: Neutralization, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Rabbit and NHP antibody responses to both prime/boost regimens were similar. New Zealand White rabbits and rhesus macaques received a priming immunization of MVAgp120 in week 0 and gp120 boosts in weeks 12 and 16. All the vaccines were either administered intramuscularly (i.m. prime/i.m. boost) or the MVAgp120 prime was administered intranasally and boosting consisted of both intramuscular and intranasal components (i.n./i.m.-plus-i.n.) ( n  = 6/group). The animals were bled at the time points indicated, and antibody responses were determined by ELISA. (A) Serum gp120-specific IgG responses after administration of the MVAgp120 prime (weeks 6 to 8), the first gp120 boost (week 15), and the second gp120 boost (week 19). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) After administration of the second booster vaccine (week 19), rabbits and NHPs had similar serum V1V2-specific IgG and serum gp120-specific IgA responses. Fecal gp120-specific responses were also similar between the species. The horizontal lines represent the geometric mean titers.
    Figure Legend Snippet: Rabbit and NHP antibody responses to both prime/boost regimens were similar. New Zealand White rabbits and rhesus macaques received a priming immunization of MVAgp120 in week 0 and gp120 boosts in weeks 12 and 16. All the vaccines were either administered intramuscularly (i.m. prime/i.m. boost) or the MVAgp120 prime was administered intranasally and boosting consisted of both intramuscular and intranasal components (i.n./i.m.-plus-i.n.) ( n  = 6/group). The animals were bled at the time points indicated, and antibody responses were determined by ELISA. (A) Serum gp120-specific IgG responses after administration of the MVAgp120 prime (weeks 6 to 8), the first gp120 boost (week 15), and the second gp120 boost (week 19). The connecting lines and error bars represent the geometric mean titers and the 95% confidence intervals. (B) After administration of the second booster vaccine (week 19), rabbits and NHPs had similar serum V1V2-specific IgG and serum gp120-specific IgA responses. Fecal gp120-specific responses were also similar between the species. The horizontal lines represent the geometric mean titers.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Rabbits developed undetectable or low mucosal antibody responses to all of the prime/boost regimens. New Zealand White rabbits received a priming immunization of MVAgp120 either i.m. or i.n. in week 0 and gp120 boosts i.m. and/or i.n. in weeks 12 and 16. Feces were collected from all individuals ( n  = 4 to 6/group), and vaginal lavage was performed on the females ( n  = 3 or 4/group). The results shown are from week 19, 3 weeks after administration of the second booster vaccine. (A) Fifty to 100% of the rabbits in each group developed detectable fecal gp120-specific IgG titers. (B) Twenty-five to 50% of the rabbits in each group developed fecal gp120-specific IgA titers. (C) Vaginal gp120-specific IgG was detectable in 50 to 100% of the female rabbits in each group but tended to be lower than the fecal gp120-specific IgG titers observed. (D) Vaginal gp120-specific IgA was detectable in only 2 of the 4 i.m./i.n.-vaccinated females. The horizontal lines indicate the geometric mean titers.
    Figure Legend Snippet: Rabbits developed undetectable or low mucosal antibody responses to all of the prime/boost regimens. New Zealand White rabbits received a priming immunization of MVAgp120 either i.m. or i.n. in week 0 and gp120 boosts i.m. and/or i.n. in weeks 12 and 16. Feces were collected from all individuals ( n  = 4 to 6/group), and vaginal lavage was performed on the females ( n  = 3 or 4/group). The results shown are from week 19, 3 weeks after administration of the second booster vaccine. (A) Fifty to 100% of the rabbits in each group developed detectable fecal gp120-specific IgG titers. (B) Twenty-five to 50% of the rabbits in each group developed fecal gp120-specific IgA titers. (C) Vaginal gp120-specific IgG was detectable in 50 to 100% of the female rabbits in each group but tended to be lower than the fecal gp120-specific IgG titers observed. (D) Vaginal gp120-specific IgA was detectable in only 2 of the 4 i.m./i.n.-vaccinated females. The horizontal lines indicate the geometric mean titers.

    Techniques Used:

    10) Product Images from "Infection of Hysterectomized Mice with Chlamydia muridarum and Chlamydia trachomatis"

    Article Title: Infection of Hysterectomized Mice with Chlamydia muridarum and Chlamydia trachomatis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00197-17

    Serum and vaginal wash ELISA antibody responses of hysterectomized mice infected with C. muridarum and C. trachomatis . (A) Serum antibody; (B) vaginal wash antibody. Vaginal infection of mice with C. muridarum resulted in a robust serum IgG2a and IgA antibody response and detectable IgG and IgA in the vaginal wash. In contrast, C. trachomatis -infected mice had no or negligible amounts of specific serum or vaginal wash antibodies ( P
    Figure Legend Snippet: Serum and vaginal wash ELISA antibody responses of hysterectomized mice infected with C. muridarum and C. trachomatis . (A) Serum antibody; (B) vaginal wash antibody. Vaginal infection of mice with C. muridarum resulted in a robust serum IgG2a and IgA antibody response and detectable IgG and IgA in the vaginal wash. In contrast, C. trachomatis -infected mice had no or negligible amounts of specific serum or vaginal wash antibodies ( P

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

    11) Product Images from "Extreme lymphoproliferative disease and fatal autoimmune thrombocytopenia in FasL and TRAIL double-deficient mice"

    Article Title: Extreme lymphoproliferative disease and fatal autoimmune thrombocytopenia in FasL and TRAIL double-deficient mice

    Journal: Blood

    doi: 10.1182/blood-2009-11-255497

    Autoantibody production in B6.GT mice . (A) Spontaneous autoimmune skin lesions and ear pinna erosion in old B6.GT mice. (B) IgM and IgG anti–double-stranded DNA-specific autoantibodies in serum from 5 age-matched female B6.WT, B6.TRAIL −/−
    Figure Legend Snippet: Autoantibody production in B6.GT mice . (A) Spontaneous autoimmune skin lesions and ear pinna erosion in old B6.GT mice. (B) IgM and IgG anti–double-stranded DNA-specific autoantibodies in serum from 5 age-matched female B6.WT, B6.TRAIL −/−

    Techniques Used: Mouse Assay

    12) Product Images from "Monoclonal antibodies detect receptor-induced binding sites in Glu-plasminogen"

    Article Title: Monoclonal antibodies detect receptor-induced binding sites in Glu-plasminogen

    Journal: Blood

    doi: 10.1182/blood-2010-11-316943

    Western blotting of anti-Pg mAbs. Glu-Pg, K1-3, K4, and K5-PD were each electrophoresed on 12.5% SDS gels under nonreducing conditions and Western blotted with the indicated mAbs (panels A-F). In controls, no bands were detected when normal mouse IgG
    Figure Legend Snippet: Western blotting of anti-Pg mAbs. Glu-Pg, K1-3, K4, and K5-PD were each electrophoresed on 12.5% SDS gels under nonreducing conditions and Western blotted with the indicated mAbs (panels A-F). In controls, no bands were detected when normal mouse IgG

    Techniques Used: Western Blot

    13) Product Images from "Thioredoxin Reductase 1 Is a Highly Immunogenic Cell Surface Antigen in Paracoccidioides spp., Candida albicans, and Cryptococcus neoformans"

    Article Title: Thioredoxin Reductase 1 Is a Highly Immunogenic Cell Surface Antigen in Paracoccidioides spp., Candida albicans, and Cryptococcus neoformans

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2019.02930

    Heterospecific and homospecific antibodies to TRR1. (A) Titers of homospecific and heterospecific antibodies in animals immunized with TRR1. Each animal was immunized three times with TRR1 from a single species or with the protein from all three species, one at a time. Two months after the last injection their sera were collected and used in ELISA experiments in which the plates were coated with TRR1 from each of the three fungal species. Bars represent the titers of antibodies (IgA + IgG + IgM) against each one of those species. (B) Sera from mice that were immunized with all three fungal species in sequence were used in western blot experiments to detect binding to recombinant C. albicans (lane b ), C. neoformans (lanes a , d ), and P. lutzii (lane e ) TRR1 proteins, as well as the protein extract of the human cell line HEK293 (lanes c , f ).
    Figure Legend Snippet: Heterospecific and homospecific antibodies to TRR1. (A) Titers of homospecific and heterospecific antibodies in animals immunized with TRR1. Each animal was immunized three times with TRR1 from a single species or with the protein from all three species, one at a time. Two months after the last injection their sera were collected and used in ELISA experiments in which the plates were coated with TRR1 from each of the three fungal species. Bars represent the titers of antibodies (IgA + IgG + IgM) against each one of those species. (B) Sera from mice that were immunized with all three fungal species in sequence were used in western blot experiments to detect binding to recombinant C. albicans (lane b ), C. neoformans (lanes a , d ), and P. lutzii (lane e ) TRR1 proteins, as well as the protein extract of the human cell line HEK293 (lanes c , f ).

    Techniques Used: Injection, Enzyme-linked Immunosorbent Assay, Mouse Assay, Sequencing, Western Blot, Binding Assay, Recombinant

    14) Product Images from "Modulation of Murine Lyme Borreliosis by Interruption of the B7/CD28 T-Cell Costimulatory Pathway"

    Article Title: Modulation of Murine Lyme Borreliosis by Interruption of the B7/CD28 T-Cell Costimulatory Pathway

    Journal: Infection and Immunity

    doi:

    Enhanced IFN-γ production by LNCs from B. burgdorferi -infected BALB/c mice treated with anti-CD80 and/or anti-CD86 MAb or CTLA-4Ig. (A) BALB/c mice were treated with antibodies to the indicated molecules or with control rat IgG as described in Materials and Methods. Bars represent the mean amount of IFN-γ present in the supernatants of pooled LNCs stimulated with B. burgdorferi lysates, as determined by ELISA, + standard error of the mean. These results are representative of three separate experiments. (B) BALB/c mice were treated with anti-CD80 and anti-CD86 MAbs, rat IgG, CTLA-4Ig, or L6 (control) as described in Materials and Methods. Bars represent the mean amount of IFN-γ produced by LNCs after restimulation with B. burgdorferi lysate, as determined by ELISA, + standard error of the mean. These results are representative of two separate experiments.
    Figure Legend Snippet: Enhanced IFN-γ production by LNCs from B. burgdorferi -infected BALB/c mice treated with anti-CD80 and/or anti-CD86 MAb or CTLA-4Ig. (A) BALB/c mice were treated with antibodies to the indicated molecules or with control rat IgG as described in Materials and Methods. Bars represent the mean amount of IFN-γ present in the supernatants of pooled LNCs stimulated with B. burgdorferi lysates, as determined by ELISA, + standard error of the mean. These results are representative of three separate experiments. (B) BALB/c mice were treated with anti-CD80 and anti-CD86 MAbs, rat IgG, CTLA-4Ig, or L6 (control) as described in Materials and Methods. Bars represent the mean amount of IFN-γ produced by LNCs after restimulation with B. burgdorferi lysate, as determined by ELISA, + standard error of the mean. These results are representative of two separate experiments.

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

    15) Product Images from "Activation of diverse repertoires of autoreactive T cells enhances the loss of anti-dsDNA B cell tolerance"

    Article Title: Activation of diverse repertoires of autoreactive T cells enhances the loss of anti-dsDNA B cell tolerance

    Journal: Journal of Clinical Investigation

    doi: 10.1172/JCI200318310

    Activation of a diverse T cell repertoire is required for the production of IgG ANAs. Activation of autoreactive CD4 + T cells leads to the expansion of autoreactive anti-dsDNA B cells that enter the B cell follicle to produce IgM ANAs. Each individual anti-dsDNA B cell requires interaction with a diverse array of T cells. T cell diversity is necessary to provide a pool of cells that can interact with the different MHC-peptide epitopes expressed on each autoreactive B cell. These multiple T cell–derived interactions are necessary for B7.2 expression on anti-dsDNA B cells; B7.2 expression on anti-dsDNA B cells may educate the T cells to migrate into the follicle. In the follicle, T cells aid autoreactive follicular B cells to produce high-affinity, isotype-switched ANAs. T, T cells; B, B cells; PALS, peri-arteriolar lymphoid sheath.
    Figure Legend Snippet: Activation of a diverse T cell repertoire is required for the production of IgG ANAs. Activation of autoreactive CD4 + T cells leads to the expansion of autoreactive anti-dsDNA B cells that enter the B cell follicle to produce IgM ANAs. Each individual anti-dsDNA B cell requires interaction with a diverse array of T cells. T cell diversity is necessary to provide a pool of cells that can interact with the different MHC-peptide epitopes expressed on each autoreactive B cell. These multiple T cell–derived interactions are necessary for B7.2 expression on anti-dsDNA B cells; B7.2 expression on anti-dsDNA B cells may educate the T cells to migrate into the follicle. In the follicle, T cells aid autoreactive follicular B cells to produce high-affinity, isotype-switched ANAs. T, T cells; B, B cells; PALS, peri-arteriolar lymphoid sheath.

    Techniques Used: Activation Assay, Derivative Assay, Expressing

    16) Product Images from "B Cell Proliferation, Somatic Hypermutation, Class Switch Recombination, and Autoantibody Production in Ectopic Lymphoid Tissue in Murine Lupus"

    Article Title: B Cell Proliferation, Somatic Hypermutation, Class Switch Recombination, and Autoantibody Production in Ectopic Lymphoid Tissue in Murine Lupus

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

    doi: 10.4049/jimmunol.0800771

    TMPD lipogranulomas contain class switched B cells A, Lipogranulomas express AID. Left , cDNA from TMPD or mineral oil lipogranulomas, spleen, or peritoneal cells was amplified using primers specific for AID or β-actin and analyzed by agarose gel electrophoresis. Right , AID mRNA was quantified by real-time PCR normalized to 18S RNA. B, μ and γ H-chain transcripts. Lipogranuloma and spleen cDNA from mineral oil or TMPD treated mice was tested for expression of IgM and IgG1 by PCR using VHF1-8 and Cμ or Cγ1 reverse primers, respectively. Left, agarose gel of amplified PCR products from lipogranulomas or spleen. γ1 transcripts are seen strongly in two TMPD lipogranulomas (TMPD, lanes 1 and 4) and weakly in another (TMPD, lane 3). Right, frequencies of IgM and IgG1 production in TMPD vs. mineral oil lipogranulomas (4 individual lipogranulomas/mouse, 3 mice/group). C, ) (arrows). PCR using B-actin primers was used as a loading control. D, Direct immunofluorescence for IgM (top) and IgG (bottom) producing cells in lipogranulomas from mineral oil or TMPD-treated mice (green). Nuclei were visualized by DAPI staining (blue). Both IgM and IgG producing cells were detected in ectopic lymphoid tissue from TMPD-treated mice, but only IgM producing cells in tissue from mineral oil-treated mice.
    Figure Legend Snippet: TMPD lipogranulomas contain class switched B cells A, Lipogranulomas express AID. Left , cDNA from TMPD or mineral oil lipogranulomas, spleen, or peritoneal cells was amplified using primers specific for AID or β-actin and analyzed by agarose gel electrophoresis. Right , AID mRNA was quantified by real-time PCR normalized to 18S RNA. B, μ and γ H-chain transcripts. Lipogranuloma and spleen cDNA from mineral oil or TMPD treated mice was tested for expression of IgM and IgG1 by PCR using VHF1-8 and Cμ or Cγ1 reverse primers, respectively. Left, agarose gel of amplified PCR products from lipogranulomas or spleen. γ1 transcripts are seen strongly in two TMPD lipogranulomas (TMPD, lanes 1 and 4) and weakly in another (TMPD, lane 3). Right, frequencies of IgM and IgG1 production in TMPD vs. mineral oil lipogranulomas (4 individual lipogranulomas/mouse, 3 mice/group). C, ) (arrows). PCR using B-actin primers was used as a loading control. D, Direct immunofluorescence for IgM (top) and IgG (bottom) producing cells in lipogranulomas from mineral oil or TMPD-treated mice (green). Nuclei were visualized by DAPI staining (blue). Both IgM and IgG producing cells were detected in ectopic lymphoid tissue from TMPD-treated mice, but only IgM producing cells in tissue from mineral oil-treated mice.

    Techniques Used: Amplification, Agarose Gel Electrophoresis, Real-time Polymerase Chain Reaction, Mouse Assay, Expressing, Polymerase Chain Reaction, Immunofluorescence, Staining

    IgG1 and IgG2a induced hypergammaglobulinemia in TMPD-treated mice is T cell dependent A, Serum samples were obtained from wild type C57BL/6J (WT, n = 5) or B6.129P2- Tcrb tm1Mom Tcrd Tm1Mom (n = 6) mice treated 3 months earlier with TMPD. IgG1, IgG2a, IgG3, and IgM levels were measured by ELISA and means were compared by the Mann-Whitney test. B, IgG production in lipogranulomas. Lipogranuloma cells and splenocytes from two mice were tested in quadruplicate for T cell dependent immunoglobulin secretion (IgG1+IgG2a+IgG2b) by ELISPOT assay. C, Quantification of plasmablasts in spleen and lipogranulomas. Pooled lipogranuloma and spleen cells from four TMPD-treated BALBc/J mice were stained with anti-B220 and anti-CD138 antibodies and analyzed by flow cytometry.
    Figure Legend Snippet: IgG1 and IgG2a induced hypergammaglobulinemia in TMPD-treated mice is T cell dependent A, Serum samples were obtained from wild type C57BL/6J (WT, n = 5) or B6.129P2- Tcrb tm1Mom Tcrd Tm1Mom (n = 6) mice treated 3 months earlier with TMPD. IgG1, IgG2a, IgG3, and IgM levels were measured by ELISA and means were compared by the Mann-Whitney test. B, IgG production in lipogranulomas. Lipogranuloma cells and splenocytes from two mice were tested in quadruplicate for T cell dependent immunoglobulin secretion (IgG1+IgG2a+IgG2b) by ELISPOT assay. C, Quantification of plasmablasts in spleen and lipogranulomas. Pooled lipogranuloma and spleen cells from four TMPD-treated BALBc/J mice were stained with anti-B220 and anti-CD138 antibodies and analyzed by flow cytometry.

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Enzyme-linked Immunospot, Staining, Flow Cytometry, Cytometry

    IgG anti-nRNP/Sm autoantibody production in TMPD-treated mice is T cell dependent A and B, Serum samples were obtained from wild type C57BL/6J (WT, n = 5) or B6.129P2- Tcrb tm1Mom Tcrd Tm1Mom (n = 6) mice treated 3 months earlier with TMPD. IgM (A) and IgG (B) anti-nRNP/Sm antibody levels were measured by ELISA at a 1:500 serum dilution. Means were compared by the Mann-Whitney test. C, Reactivity of sera with recombinant U1-A protein (ELISA). Recombinant 6His-tagged U1-A protein was expressed in E. coli and purified on a Ni-NTA affinity column. Sera from 20 TMPD-treated BALB/c mice positive for anti-Sm/RNP autoantibodies and 20 normal BALB/c mouse sera were tested for reactivity with the recombinant antigen at a 1:100 dilution (ELISA). D, IgM and IgG ELISPOT assay with purified U1-A antigen using cells isolated from collagenase treated ectopic lymphoid tissue from an anti-RNP positive TMPD-treated mouse or an anti-RNP negative mouse treated with medicinal mineral oil. Representative of 3 experiments. E, IgG ELISPOT assay with purified U1-A or bovine serum albumin (BSA) antigens, using cells isolated from lipogranulomas (Lipogran) or spleens of anti-U1-A positive mice (n = 5). The frequencies of antigen-specific spots are expressed per 50,000 B cells. The frequency of anti-U1-A spots was higher in lipogranulomas than in spleen (P = 0.01, Mann-Whitney test) and the frequency of anti-U1-A spots was higher than the frequency of anti-BSA spots (P = 0.03, Mann-Whitney test).
    Figure Legend Snippet: IgG anti-nRNP/Sm autoantibody production in TMPD-treated mice is T cell dependent A and B, Serum samples were obtained from wild type C57BL/6J (WT, n = 5) or B6.129P2- Tcrb tm1Mom Tcrd Tm1Mom (n = 6) mice treated 3 months earlier with TMPD. IgM (A) and IgG (B) anti-nRNP/Sm antibody levels were measured by ELISA at a 1:500 serum dilution. Means were compared by the Mann-Whitney test. C, Reactivity of sera with recombinant U1-A protein (ELISA). Recombinant 6His-tagged U1-A protein was expressed in E. coli and purified on a Ni-NTA affinity column. Sera from 20 TMPD-treated BALB/c mice positive for anti-Sm/RNP autoantibodies and 20 normal BALB/c mouse sera were tested for reactivity with the recombinant antigen at a 1:100 dilution (ELISA). D, IgM and IgG ELISPOT assay with purified U1-A antigen using cells isolated from collagenase treated ectopic lymphoid tissue from an anti-RNP positive TMPD-treated mouse or an anti-RNP negative mouse treated with medicinal mineral oil. Representative of 3 experiments. E, IgG ELISPOT assay with purified U1-A or bovine serum albumin (BSA) antigens, using cells isolated from lipogranulomas (Lipogran) or spleens of anti-U1-A positive mice (n = 5). The frequencies of antigen-specific spots are expressed per 50,000 B cells. The frequency of anti-U1-A spots was higher in lipogranulomas than in spleen (P = 0.01, Mann-Whitney test) and the frequency of anti-U1-A spots was higher than the frequency of anti-BSA spots (P = 0.03, Mann-Whitney test).

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Recombinant, Purification, Affinity Column, Enzyme-linked Immunospot, Isolation

    17) Product Images from "Toll-like receptor 8 deletion accelerates autoimmunity in a mouse model of lupus through a Toll-like receptor 7-dependent mechanism"

    Article Title: Toll-like receptor 8 deletion accelerates autoimmunity in a mouse model of lupus through a Toll-like receptor 7-dependent mechanism

    Journal: Immunology

    doi: 10.1111/imm.12426

    Anti-dsDNA and antinuclear autoantibodies are detected in male Nba2. Yaa and Nba2.TLR8 − / Yaa . Staining of Crithidia luciliae or Hep2 cell coated slides for the presence of IgG anti-dsDNA or IgG antinuclear autoantibodies in the serum of Nba2. Yaa
    Figure Legend Snippet: Anti-dsDNA and antinuclear autoantibodies are detected in male Nba2. Yaa and Nba2.TLR8 − / Yaa . Staining of Crithidia luciliae or Hep2 cell coated slides for the presence of IgG anti-dsDNA or IgG antinuclear autoantibodies in the serum of Nba2. Yaa

    Techniques Used: Staining

    18) Product Images from "Gammaherpesvirus Co-infection with Malaria Suppresses Anti-parasitic Humoral Immunity"

    Article Title: Gammaherpesvirus Co-infection with Malaria Suppresses Anti-parasitic Humoral Immunity

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1004858

    MHV68 co-infection with the non-lethal P . yoelii XNL in C57BL/6 results in lethal malarial disease and suppressed Plasmodium specific IgG response. (A) Timeline of infection. 6–8 week old C57BL/6 mice were infected with 1000 PFU of MHV68 on day -7 followed by infection with 10 5 pRBCs of non-lethal P . yoelii XNL or P . chabaudi AS. Infections consisted of 5 experimental groups: MHV68 + Plasmodium , Plasmodium , MHV68 or mock infected. Each experimental group consisted of n = 5 and was repeated twice. Animals were sacrificed at days 8, 12, 16 and 23 post P . yoelii XNL infection or day 7, 11, 15 and 23 post P . chabaudi AS infection for collection of spleen, lung and blood. (B) Survival analysis of animals co-infected with MHV68 and P . yoelii XNL or P . chabaudi AS. Total IgG and IgM levels in serum in (C) P . yoelii XNL (Day 23 IgG— P . yoelii vs co-infected: p
    Figure Legend Snippet: MHV68 co-infection with the non-lethal P . yoelii XNL in C57BL/6 results in lethal malarial disease and suppressed Plasmodium specific IgG response. (A) Timeline of infection. 6–8 week old C57BL/6 mice were infected with 1000 PFU of MHV68 on day -7 followed by infection with 10 5 pRBCs of non-lethal P . yoelii XNL or P . chabaudi AS. Infections consisted of 5 experimental groups: MHV68 + Plasmodium , Plasmodium , MHV68 or mock infected. Each experimental group consisted of n = 5 and was repeated twice. Animals were sacrificed at days 8, 12, 16 and 23 post P . yoelii XNL infection or day 7, 11, 15 and 23 post P . chabaudi AS infection for collection of spleen, lung and blood. (B) Survival analysis of animals co-infected with MHV68 and P . yoelii XNL or P . chabaudi AS. Total IgG and IgM levels in serum in (C) P . yoelii XNL (Day 23 IgG— P . yoelii vs co-infected: p

    Techniques Used: Infection, Mouse Assay

    19) Product Images from "Immunization with Ehrlichia P28 Outer Membrane Proteins Confers Protection in a Mouse Model of Ehrlichiosis ▿"

    Article Title: Immunization with Ehrlichia P28 Outer Membrane Proteins Confers Protection in a Mouse Model of Ehrlichiosis ▿

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.05292-11

    Induction of Ehrlichia -specific IgG1, IgG2b, and IgG3 antibody isotypes was associated with protection in mice immunized with P28-19 protein. P28-19-specific antibody responses in mice immunized with the P28-19 protein on days 7, 14, and 21 after E. muris challenge were measured by ELISAs. (A) Comparable concentrations of P28-19-specific IgM and IgG2b antibodies were found in sera from mice immunized with P28-19 protein and unimmunized mice on day 7 after E. muris challenge. (B) Mice immunized with the recombinant P28-19 protein had higher concentrations of P28-19-specific IgG1, IgG2b, and IgG3 antibodies on day 14 after E. muris challenge than the controls. (C) In contrast, unimmunized mice challenged with E. muris developed P28-19-specific IgG2c, IgG2b, and IgG3 antibodies by day 21 postchallenge. The data were expressed as means plus standard deviations, and three mice from each group were used for this analysis.
    Figure Legend Snippet: Induction of Ehrlichia -specific IgG1, IgG2b, and IgG3 antibody isotypes was associated with protection in mice immunized with P28-19 protein. P28-19-specific antibody responses in mice immunized with the P28-19 protein on days 7, 14, and 21 after E. muris challenge were measured by ELISAs. (A) Comparable concentrations of P28-19-specific IgM and IgG2b antibodies were found in sera from mice immunized with P28-19 protein and unimmunized mice on day 7 after E. muris challenge. (B) Mice immunized with the recombinant P28-19 protein had higher concentrations of P28-19-specific IgG1, IgG2b, and IgG3 antibodies on day 14 after E. muris challenge than the controls. (C) In contrast, unimmunized mice challenged with E. muris developed P28-19-specific IgG2c, IgG2b, and IgG3 antibodies by day 21 postchallenge. The data were expressed as means plus standard deviations, and three mice from each group were used for this analysis.

    Techniques Used: Mouse Assay, Recombinant

    Protection induced by the recombinant P28-19 protein was associated with the development of antigen-specific IgG responses. P28-19-specific antibody responses were measured by an ELISA using a 22-amino-acid synthetic P28-19 peptide as the antigen. Mice immunized with the recombinant E. muris P28-19 protein had higher concentrations of P28-19-specific IgG antibodies on day 14 after E. muris challenge than mice in the unvaccinated control group. Antibody responses in mice immunized with the recombinant P28-19 alone are presented for comparison. The data were expressed as means plus standard deviations, and three mice from each group were used for this analysis. OD650nm, optical density at 650 nm.
    Figure Legend Snippet: Protection induced by the recombinant P28-19 protein was associated with the development of antigen-specific IgG responses. P28-19-specific antibody responses were measured by an ELISA using a 22-amino-acid synthetic P28-19 peptide as the antigen. Mice immunized with the recombinant E. muris P28-19 protein had higher concentrations of P28-19-specific IgG antibodies on day 14 after E. muris challenge than mice in the unvaccinated control group. Antibody responses in mice immunized with the recombinant P28-19 alone are presented for comparison. The data were expressed as means plus standard deviations, and three mice from each group were used for this analysis. OD650nm, optical density at 650 nm.

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

    Ehrlichia -specific IgG antibody responses develop in mice immunized with the P28 outer membrane proteins. Ehrlichia muris -specific IgG antibody titers in all vaccinated groups before and after challenge were measured by IFA using slides prepared from E. muris cultured in DH82 cells. Higher titers of E. muris -specific IgG antibodies were observed in mice vaccinated with P28-9, P28-12, P28-14, P28-19, and the P28 mixture on day 28 after the last booster immunization before challenge than in the controls. The titers of E. muris -specific IgG antibodies measured after challenge were higher than the titers of antibodies in vaccinated mice before challenge, indicating that infection boosted the antibody response. Each symbol shows the value for an individual mouse. The short horizontal lines represent the means for the groups of mice before challenge (short broken lines) and after challenge (short black lines).
    Figure Legend Snippet: Ehrlichia -specific IgG antibody responses develop in mice immunized with the P28 outer membrane proteins. Ehrlichia muris -specific IgG antibody titers in all vaccinated groups before and after challenge were measured by IFA using slides prepared from E. muris cultured in DH82 cells. Higher titers of E. muris -specific IgG antibodies were observed in mice vaccinated with P28-9, P28-12, P28-14, P28-19, and the P28 mixture on day 28 after the last booster immunization before challenge than in the controls. The titers of E. muris -specific IgG antibodies measured after challenge were higher than the titers of antibodies in vaccinated mice before challenge, indicating that infection boosted the antibody response. Each symbol shows the value for an individual mouse. The short horizontal lines represent the means for the groups of mice before challenge (short broken lines) and after challenge (short black lines).

    Techniques Used: Mouse Assay, Immunofluorescence, Cell Culture, Infection

    20) Product Images from "IRAK-M Deficiency Promotes the Development of Type 1 Diabetes in NOD Mice"

    Article Title: IRAK-M Deficiency Promotes the Development of Type 1 Diabetes in NOD Mice

    Journal: Diabetes

    doi: 10.2337/db13-1504

    IRAK-M deficiency promotes the development of T1DM in NOD mice. A : Diabetes incidence in female IRAK-M −/− NOD and IRAK-M +/− NOD mice, and their WT NOD littermates. Statistical analysis was performed by Gehan-Breslow-Wilcoxon test. B : An intraperitoneal glucose tolerance test was performed in ∼12-week-old female IRAK-M −/− NOD and WT NOD mice that were fasted overnight with free access to water. Blood glucose was measured at the indicated time points after glucose injection (2 g/kg body weight i.p.; n = 5 mice/group), and data were analyzed by two-way ANOVA. C : Serum anti-insulin IgG was measured in serum samples of 12-week-old nondiabetic female IRAK-M −/− NOD and WT NOD mice by ELISA ( n = 24/group). Data are presented as an optical density (OD) of 405 nm and were analyzed by Student t test. D : Histology of islet infiltration in 12-week-old nondiabetic female IRAK-M −/− NOD and WT NOD mice. Hematoxylin-eosin staining of the pancreas showed more immune cell infiltration in islets of IRAK-M −/− NOD mice compared with the control mice (top). Insulitis was quantified using the following scoring system: 0, no insulitis; 1,
    Figure Legend Snippet: IRAK-M deficiency promotes the development of T1DM in NOD mice. A : Diabetes incidence in female IRAK-M −/− NOD and IRAK-M +/− NOD mice, and their WT NOD littermates. Statistical analysis was performed by Gehan-Breslow-Wilcoxon test. B : An intraperitoneal glucose tolerance test was performed in ∼12-week-old female IRAK-M −/− NOD and WT NOD mice that were fasted overnight with free access to water. Blood glucose was measured at the indicated time points after glucose injection (2 g/kg body weight i.p.; n = 5 mice/group), and data were analyzed by two-way ANOVA. C : Serum anti-insulin IgG was measured in serum samples of 12-week-old nondiabetic female IRAK-M −/− NOD and WT NOD mice by ELISA ( n = 24/group). Data are presented as an optical density (OD) of 405 nm and were analyzed by Student t test. D : Histology of islet infiltration in 12-week-old nondiabetic female IRAK-M −/− NOD and WT NOD mice. Hematoxylin-eosin staining of the pancreas showed more immune cell infiltration in islets of IRAK-M −/− NOD mice compared with the control mice (top). Insulitis was quantified using the following scoring system: 0, no insulitis; 1,

    Techniques Used: Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay, Staining

    21) Product Images from "Suppression of lupus nephritis and skin lesions in MRL/lpr mice by administration of the topoisomerase I inhibitor irinotecan"

    Article Title: Suppression of lupus nephritis and skin lesions in MRL/lpr mice by administration of the topoisomerase I inhibitor irinotecan

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-016-1144-5

    Increased binding of anti-double-stranded DNS ( dsDNA ) IgG but not of anti-dsDNA IgM to DNA treated with topoisomerase I ( topo I ). Calf thymus DNA was incubated with recombinant topo I or bovine serum albumin ( BSA ) for 30 min. Binding affinity of a anti-dsDNA IgG or b anti-dsDNA IgM from MRL/ lpr plasma to modified DNA was assessed by enzyme-linked immunosorbent assay. Data were pooled from three independent experiments. ** P
    Figure Legend Snippet: Increased binding of anti-double-stranded DNS ( dsDNA ) IgG but not of anti-dsDNA IgM to DNA treated with topoisomerase I ( topo I ). Calf thymus DNA was incubated with recombinant topo I or bovine serum albumin ( BSA ) for 30 min. Binding affinity of a anti-dsDNA IgG or b anti-dsDNA IgM from MRL/ lpr plasma to modified DNA was assessed by enzyme-linked immunosorbent assay. Data were pooled from three independent experiments. ** P

    Techniques Used: Binding Assay, Incubation, Recombinant, Modification, Enzyme-linked Immunosorbent Assay

    Decreased anti-double-stranded DNA ( dsDNA ) IgM in MRL/ lpr mice treated with low-dose irinotecan ( Irino ). MRL/ lpr mice receiving saline ( n = 7), 25 mg/kg irinotecan ( n = 6), or 1 mg/kg irinotecan ( n = 6) were sacrificed at 18 weeks of age. MRL/MpJ mice treated with saline ( n = 7) were used as controls ( Ctrl ). a Number of IgG- and IgM-secreting cells in the spleen determined by enzyme-linked immunosorbent spot assay. b Plasma levels of total IgG and IgM and c anti-dsDNA-specific IgG and IgM were measured by enzyme-linked immunosorbent assay. In ( a ), each symbol represents an individual mouse; bars show mean ± SEM. * P
    Figure Legend Snippet: Decreased anti-double-stranded DNA ( dsDNA ) IgM in MRL/ lpr mice treated with low-dose irinotecan ( Irino ). MRL/ lpr mice receiving saline ( n = 7), 25 mg/kg irinotecan ( n = 6), or 1 mg/kg irinotecan ( n = 6) were sacrificed at 18 weeks of age. MRL/MpJ mice treated with saline ( n = 7) were used as controls ( Ctrl ). a Number of IgG- and IgM-secreting cells in the spleen determined by enzyme-linked immunosorbent spot assay. b Plasma levels of total IgG and IgM and c anti-dsDNA-specific IgG and IgM were measured by enzyme-linked immunosorbent assay. In ( a ), each symbol represents an individual mouse; bars show mean ± SEM. * P

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

    22) Product Images from "CTLA4Ig Prevents Initiation but not Evolution of Anti-phospholipid Syndrome in NZW/BXSB Mice"

    Article Title: CTLA4Ig Prevents Initiation but not Evolution of Anti-phospholipid Syndrome in NZW/BXSB Mice

    Journal: Autoimmunity

    doi: 10.1080/08916930400008524

    Frequency of Ig secreting cells analyzed by ELISpot. Mean + SD frequency of antibody and autoantibody secreting B cells of the IgM and IgG isotypes from spleens of untreated and treated mice. Spleens from untreated and 12 week treated mice were analyzed at 22 weeks. Frequency is per 10 3 cells for total Ig and per 10 5 cells for anti-dsDNA Ig and anti-cardiolipin Ig. Mice treated with CTLA4Ig at 12 weeks were not different from untreated mice. Mice treated with CTLA4Ig at 9 weeks had a significantly lower frequency of Ig and autoantibody-secreting cells than untreated controls ( p
    Figure Legend Snippet: Frequency of Ig secreting cells analyzed by ELISpot. Mean + SD frequency of antibody and autoantibody secreting B cells of the IgM and IgG isotypes from spleens of untreated and treated mice. Spleens from untreated and 12 week treated mice were analyzed at 22 weeks. Frequency is per 10 3 cells for total Ig and per 10 5 cells for anti-dsDNA Ig and anti-cardiolipin Ig. Mice treated with CTLA4Ig at 12 weeks were not different from untreated mice. Mice treated with CTLA4Ig at 9 weeks had a significantly lower frequency of Ig and autoantibody-secreting cells than untreated controls ( p

    Techniques Used: Enzyme-linked Immunospot, Mouse Assay

    23) Product Images from "Conjugation of Mannans to Enhance the Potency of Liposome Nanoparticles for the Delivery of RNA Vaccines"

    Article Title: Conjugation of Mannans to Enhance the Potency of Liposome Nanoparticles for the Delivery of RNA Vaccines

    Journal: Pharmaceutics

    doi: 10.3390/pharmaceutics13020240

    Immunization titers of RSV F SAM antigen formulated with classical 2% PEG LNP and MLNP formulations and administered through IM route. Median IgG ( A ), IgG1 ( B ), and IgG2a ( C ) titers, considering non adjuvanted subunit (SU) group as control. * p
    Figure Legend Snippet: Immunization titers of RSV F SAM antigen formulated with classical 2% PEG LNP and MLNP formulations and administered through IM route. Median IgG ( A ), IgG1 ( B ), and IgG2a ( C ) titers, considering non adjuvanted subunit (SU) group as control. * p

    Techniques Used:

    Immunization titers of RSV F SAM antigen formulated with classical 2% PEG LNP and MLNP formulations and administered through ID route. Median IgG ( A ), IgG1 ( B ), and IgG2a ( C ) titers, considering non adjuvanted subunit (SU) group as control. * p
    Figure Legend Snippet: Immunization titers of RSV F SAM antigen formulated with classical 2% PEG LNP and MLNP formulations and administered through ID route. Median IgG ( A ), IgG1 ( B ), and IgG2a ( C ) titers, considering non adjuvanted subunit (SU) group as control. * p

    Techniques Used:

    Immunization titers of RSV F SAM antigen formulated with classical 0.3% PEG LNP and MLNP formulations and administered through ID route. Median IgG ( A ), IgG1 ( B ), and IgG2a ( C ) titers, considering non adjuvanted subunit (SU) group as control. * p
    Figure Legend Snippet: Immunization titers of RSV F SAM antigen formulated with classical 0.3% PEG LNP and MLNP formulations and administered through ID route. Median IgG ( A ), IgG1 ( B ), and IgG2a ( C ) titers, considering non adjuvanted subunit (SU) group as control. * p

    Techniques Used:

    24) Product Images from "Robust IgA and IgG-producing antibody forming cells in the diffuse NALT and lungs of Sendai virus-vaccinated cotton rats associate with rapid protection against human parainfluenza virus-type 1"

    Article Title: Robust IgA and IgG-producing antibody forming cells in the diffuse NALT and lungs of Sendai virus-vaccinated cotton rats associate with rapid protection against human parainfluenza virus-type 1

    Journal: Vaccine

    doi: 10.1016/j.vaccine.2010.07.068

    IgA and IgG in the nasal wash of SeV-primed animals
    Figure Legend Snippet: IgA and IgG in the nasal wash of SeV-primed animals

    Techniques Used:

    Sustained IgA and IgG AFC in the d-NALT
    Figure Legend Snippet: Sustained IgA and IgG AFC in the d-NALT

    Techniques Used:

    IgA and IgG in the LRT
    Figure Legend Snippet: IgA and IgG in the LRT

    Techniques Used:

    Long-sustained virus-specific IgG, but not IgA in the serum of SeV-primed cotton rats
    Figure Legend Snippet: Long-sustained virus-specific IgG, but not IgA in the serum of SeV-primed cotton rats

    Techniques Used:

    25) Product Images from "Amplified B Lymphocyte CD40 Signaling Drives Regulatory B10 Cell Expansion in Mice"

    Article Title: Amplified B Lymphocyte CD40 Signaling Drives Regulatory B10 Cell Expansion in Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0022464

    Impaired IgG and GC responses in CD154 TG CD22 −/− mice. ( A ) Serum IgM and IgG levels of 4 and 12 mo-old WT, CD22 −/− , CD154 TG , and CD154 TG CD22 −/− mice. Symbols represent serum concentrations for individual mice as determined by ELISA, with means indicated by horizontal bars. ( B ) Serum autoAbs reactive with dsDNA, ssDNA, or histone proteins in 12 mo-old mice. ELISA OD values for IgM (upper panels) and IgG (lower panels) autoAbs are shown for individual mice, with means indicated by horizontal bars. Sera from 2 mo-old WT C57BL/6 and 6 mo-old MLR lpr mice were used as negative and positive controls, respectively. ( C ) Impaired IgG responses to a TD Ag. WT ( n = 3), CD22 −/− ( n = 4), CD154 TG ( n = 6), and CD154 TG CD22 −/− ( n = 8) mice were immunized with DNP-KLH in adjuvant on day 0, and boosted on day 21. The graph shows mean (±SEM) DNP-specific IgG levels as determined by ELISA. Images on the right represent immunofluorescence staining of frozen spleen sections from all genotypes harvested 7 days after the boost phase of DNP-KLH immunization. Merged images show the presence of B220 + B cells (red) and GC GL7 + B220 + B cells (yellow). Enlarged regions from these sections indicate typical GC structures present within the follicles of WT and CD22 −/− mice, and detectable GL7 + B220 + B cells within the follicles of CD154 TG mice, but not in CD154 TG CD22 −/− mice (representative regions are shown for comparison). (A–C) Means significantly different from WT are indicated by asterisks (*p≤0.05, **p≤0.01), and between other indicated groups by crosses (†p
    Figure Legend Snippet: Impaired IgG and GC responses in CD154 TG CD22 −/− mice. ( A ) Serum IgM and IgG levels of 4 and 12 mo-old WT, CD22 −/− , CD154 TG , and CD154 TG CD22 −/− mice. Symbols represent serum concentrations for individual mice as determined by ELISA, with means indicated by horizontal bars. ( B ) Serum autoAbs reactive with dsDNA, ssDNA, or histone proteins in 12 mo-old mice. ELISA OD values for IgM (upper panels) and IgG (lower panels) autoAbs are shown for individual mice, with means indicated by horizontal bars. Sera from 2 mo-old WT C57BL/6 and 6 mo-old MLR lpr mice were used as negative and positive controls, respectively. ( C ) Impaired IgG responses to a TD Ag. WT ( n = 3), CD22 −/− ( n = 4), CD154 TG ( n = 6), and CD154 TG CD22 −/− ( n = 8) mice were immunized with DNP-KLH in adjuvant on day 0, and boosted on day 21. The graph shows mean (±SEM) DNP-specific IgG levels as determined by ELISA. Images on the right represent immunofluorescence staining of frozen spleen sections from all genotypes harvested 7 days after the boost phase of DNP-KLH immunization. Merged images show the presence of B220 + B cells (red) and GC GL7 + B220 + B cells (yellow). Enlarged regions from these sections indicate typical GC structures present within the follicles of WT and CD22 −/− mice, and detectable GL7 + B220 + B cells within the follicles of CD154 TG mice, but not in CD154 TG CD22 −/− mice (representative regions are shown for comparison). (A–C) Means significantly different from WT are indicated by asterisks (*p≤0.05, **p≤0.01), and between other indicated groups by crosses (†p

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

    26) Product Images from "A chimeric influenza hemagglutinin delivered by parainfluenza virus 5 vector induces broadly protective immunity against genetically divergent influenza A H1 viruses in swine"

    Article Title: A chimeric influenza hemagglutinin delivered by parainfluenza virus 5 vector induces broadly protective immunity against genetically divergent influenza A H1 viruses in swine

    Journal: Veterinary Microbiology

    doi: 10.1016/j.vetmic.2020.108859

    Evaluation of PIV5-113-induced antibodies against parental HA, using ELISA: IgG (H + L), IgG1, and IgG2a. Mice were vaccinated with either PBS, PIV5, or PIV5-113, and sera were collected after delivery of a boost, as described above. Results were obtained using sera from PBS-inoculated mice (IgG (H + L): n = 18 for NJ76 and IA06, n = 20 for OH07, ME08, and TN09, IgG1: n = 18 for NJ76, IA06, OH07, ME08, and TN09, IgG2a: n = 18 for NJ76, IA06, OH07, ME08, and TN09), PIV5-inoculated mice (n = 40), and PIV5-113 (n = 38). *Indicates a significant difference (P
    Figure Legend Snippet: Evaluation of PIV5-113-induced antibodies against parental HA, using ELISA: IgG (H + L), IgG1, and IgG2a. Mice were vaccinated with either PBS, PIV5, or PIV5-113, and sera were collected after delivery of a boost, as described above. Results were obtained using sera from PBS-inoculated mice (IgG (H + L): n = 18 for NJ76 and IA06, n = 20 for OH07, ME08, and TN09, IgG1: n = 18 for NJ76, IA06, OH07, ME08, and TN09, IgG2a: n = 18 for NJ76, IA06, OH07, ME08, and TN09), PIV5-inoculated mice (n = 40), and PIV5-113 (n = 38). *Indicates a significant difference (P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay

    27) Product Images from "Toll-like receptor 9 controls anti-DNA autoantibody production in murine lupus"

    Article Title: Toll-like receptor 9 controls anti-DNA autoantibody production in murine lupus

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20050338

    Glomerular immune deposits do not require anti-DNA antibodies. (A) Glomerular immune deposits were detected by direct immunofluorescence for IgG (top) and complement C3 (bottom) in frozen kidney sections from TLR9 +/+ (left), TLR9 −/− (middle), and nonautoimmune C57BL/6 control mice (right). Representative images are shown. Original magnification, 400. (B) Mean glomerular fluorescence intensity (arbitrary units) was determined for IgG and C3 in TLR9 +/+ ( n = 8) and TLR9 −/− ( n = 8) mice. Horizontal lines represent mean values.
    Figure Legend Snippet: Glomerular immune deposits do not require anti-DNA antibodies. (A) Glomerular immune deposits were detected by direct immunofluorescence for IgG (top) and complement C3 (bottom) in frozen kidney sections from TLR9 +/+ (left), TLR9 −/− (middle), and nonautoimmune C57BL/6 control mice (right). Representative images are shown. Original magnification, 400. (B) Mean glomerular fluorescence intensity (arbitrary units) was determined for IgG and C3 in TLR9 +/+ ( n = 8) and TLR9 −/− ( n = 8) mice. Horizontal lines represent mean values.

    Techniques Used: Immunofluorescence, Mouse Assay, Fluorescence

    Lymphadenopathy, hypergammaglobulinemia, and lymphocyte accumulation in TLR-deficient mice. TLR9 +/+ ( n = 19), TLR9 −/− ( n = 16), TLR3 +/+ ( n = 15), and TLR3 −/− ( n = 17) mice were killed at 20 wk of age and assessed for evidence of aberrant immune activation; nonautoimmune C57BL/6 control mice ( n = 4) were killed at 26 wk of age. (A) Spleens and the two largest axillary lymph nodes were removed and weighed. (B) Total serum IgG and IgG2a were determined. (C) Splenocyte subsets were enumerated by FACS analysis for T cells (Thy1.2 + ), DNTC (CD4 − /CD8 − double-negative T cells), and B cells (CD22 + ). (D) Splenic CD4 + T cells were classified as either naive (CD44 − CD62L + ), activated (CD44 + CD62L + ), or memory (CD44 + CD62L − ) phenotype. The analysis in D was performed on 12 TLR3 +/+ and 12 TLR3 −/− mice. Horizontal lines represent mean values.
    Figure Legend Snippet: Lymphadenopathy, hypergammaglobulinemia, and lymphocyte accumulation in TLR-deficient mice. TLR9 +/+ ( n = 19), TLR9 −/− ( n = 16), TLR3 +/+ ( n = 15), and TLR3 −/− ( n = 17) mice were killed at 20 wk of age and assessed for evidence of aberrant immune activation; nonautoimmune C57BL/6 control mice ( n = 4) were killed at 26 wk of age. (A) Spleens and the two largest axillary lymph nodes were removed and weighed. (B) Total serum IgG and IgG2a were determined. (C) Splenocyte subsets were enumerated by FACS analysis for T cells (Thy1.2 + ), DNTC (CD4 − /CD8 − double-negative T cells), and B cells (CD22 + ). (D) Splenic CD4 + T cells were classified as either naive (CD44 − CD62L + ), activated (CD44 + CD62L + ), or memory (CD44 + CD62L − ) phenotype. The analysis in D was performed on 12 TLR3 +/+ and 12 TLR3 −/− mice. Horizontal lines represent mean values.

    Techniques Used: Mouse Assay, Activation Assay, FACS

    Reduced anti-dsDNA autoantibodies in TLR9 − / − but not TLR3 − / − mice. (A) Anti-dsDNA antibodies were detected by C. luciliae immunofluorescence. IgG antibodies to C. luciliae DNA are shown in green (left), and DAPI staining of DNA is shown in red (middle). White arrows indicate the kinetoplast. Specific anti-dsDNA antibodies are identified by colocalization of IgG and DAPI staining in the kinetoplast and appear in yellow (right). Representative TLR9 +/+ sera are shown in the top two rows (intensity scores of 3 + and 1 + ), and TLR9 −/− sera are shown in the bottom two rows (intensity scores of 1 + and 0). Original magnification, 1,000. (B and C) Specific anti-dsDNA staining of C. luciliae kinetoplasts was scored from 0 to 4 as in A for either TLR9 +/+ ( n = 19) and TLR9 −/− ( n = 16) sera (B), or TLR3 +/+ ( n = 15) and TLR3 −/− ( n = 17) sera (C). *, P
    Figure Legend Snippet: Reduced anti-dsDNA autoantibodies in TLR9 − / − but not TLR3 − / − mice. (A) Anti-dsDNA antibodies were detected by C. luciliae immunofluorescence. IgG antibodies to C. luciliae DNA are shown in green (left), and DAPI staining of DNA is shown in red (middle). White arrows indicate the kinetoplast. Specific anti-dsDNA antibodies are identified by colocalization of IgG and DAPI staining in the kinetoplast and appear in yellow (right). Representative TLR9 +/+ sera are shown in the top two rows (intensity scores of 3 + and 1 + ), and TLR9 −/− sera are shown in the bottom two rows (intensity scores of 1 + and 0). Original magnification, 1,000. (B and C) Specific anti-dsDNA staining of C. luciliae kinetoplasts was scored from 0 to 4 as in A for either TLR9 +/+ ( n = 19) and TLR9 −/− ( n = 16) sera (B), or TLR3 +/+ ( n = 15) and TLR3 −/− ( n = 17) sera (C). *, P

    Techniques Used: Mouse Assay, Immunofluorescence, Staining

    28) Product Images from "IgG3 deficiency extends lifespan and attenuates progression of glomerulonephritis in MRL/lpr mice"

    Article Title: IgG3 deficiency extends lifespan and attenuates progression of glomerulonephritis in MRL/lpr mice

    Journal: Biology Direct

    doi: 10.1186/1745-6150-7-3

    Western blot for γ3 heavy chain (IgG3) in serum from mice bearing zero (-/-), one (+/-), or two (+/+) wild-type (functional) γ3 heavy chain alleles by PCR genotyping assay . Lane 1, markers; 2-6, γ3 +/+; 7-8, γ3 +/-; 9-10, γ3 -/-.
    Figure Legend Snippet: Western blot for γ3 heavy chain (IgG3) in serum from mice bearing zero (-/-), one (+/-), or two (+/+) wild-type (functional) γ3 heavy chain alleles by PCR genotyping assay . Lane 1, markers; 2-6, γ3 +/+; 7-8, γ3 +/-; 9-10, γ3 -/-.

    Techniques Used: Western Blot, Mouse Assay, Functional Assay, Polymerase Chain Reaction, Genotyping Assay

    Serum auto-antibody activity for dsDNA or α-actinin assessed by ELISA . Serum samples from γ3 +/+ (green), +/- (yellow), and -/- (red) mice were tested for binding of total IgG (left panel) to double stranded (ds) DNA (circles) or to α-actinin (squares), and for binding of IgG3 (right panel) to dsDNA (diamonds) or to α-actinin (triangles).
    Figure Legend Snippet: Serum auto-antibody activity for dsDNA or α-actinin assessed by ELISA . Serum samples from γ3 +/+ (green), +/- (yellow), and -/- (red) mice were tested for binding of total IgG (left panel) to double stranded (ds) DNA (circles) or to α-actinin (squares), and for binding of IgG3 (right panel) to dsDNA (diamonds) or to α-actinin (triangles).

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

    29) Product Images from "Intradermal immunisation using the TLR3-ligand Poly (I:C) as adjuvant induces mucosal antibody responses and protects against genital HSV-2 infection"

    Article Title: Intradermal immunisation using the TLR3-ligand Poly (I:C) as adjuvant induces mucosal antibody responses and protects against genital HSV-2 infection

    Journal: NPJ Vaccines

    doi: 10.1038/npjvaccines.2016.10

    Ab-producing cells induced by ID immunisation with gp140+Poly(I:C). IgG ( a ) and IgA ( b ) SFC were measured by ELISPOT assay 5 days after the last ID immunisation with gp140+Poly(I:C) in uterus, colon, small intestine and bone marrow (left panels) and in cLN, mLN, gLN and spleen (right panels). Data are expressed as mean+s.e.m. of gp140-specific SFC/10 6 total ( right panels ) or CD45 + cells ( left panels ) pooled from 6–8 mice /group. Statsitical analyses were performed using one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test.
    Figure Legend Snippet: Ab-producing cells induced by ID immunisation with gp140+Poly(I:C). IgG ( a ) and IgA ( b ) SFC were measured by ELISPOT assay 5 days after the last ID immunisation with gp140+Poly(I:C) in uterus, colon, small intestine and bone marrow (left panels) and in cLN, mLN, gLN and spleen (right panels). Data are expressed as mean+s.e.m. of gp140-specific SFC/10 6 total ( right panels ) or CD45 + cells ( left panels ) pooled from 6–8 mice /group. Statsitical analyses were performed using one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test.

    Techniques Used: Enzyme-linked Immunospot, Mouse Assay

    HIV-1 gp140-specific IgG and IgA responses induced via ID route compared with those induced via s.c. and IN routes. Gp140-specific IgG (top panels) and IgA (bottom panels) were titrated in serum ( a ), vaginal fluids ( b ) and fecal extracts ( c ) of mice at various time points after the last immunisation with gp140 plus Poly(I:C) via either the ID (black squares), s.c. (white squares) or IN (black triangles) route. Results are expressed as mean+s.e.m. of Ab titres and are representative of one out of two experiments using 6 mice per group. Statistical analyses were performed using the two-way analysis of variance (ANOVA) test and Bonferroni’s multiple comparisons.
    Figure Legend Snippet: HIV-1 gp140-specific IgG and IgA responses induced via ID route compared with those induced via s.c. and IN routes. Gp140-specific IgG (top panels) and IgA (bottom panels) were titrated in serum ( a ), vaginal fluids ( b ) and fecal extracts ( c ) of mice at various time points after the last immunisation with gp140 plus Poly(I:C) via either the ID (black squares), s.c. (white squares) or IN (black triangles) route. Results are expressed as mean+s.e.m. of Ab titres and are representative of one out of two experiments using 6 mice per group. Statistical analyses were performed using the two-way analysis of variance (ANOVA) test and Bonferroni’s multiple comparisons.

    Techniques Used: Mouse Assay

    Ampligen promotes HIV-1 gp140-specific mucosal IgG but not IgA. Mice were immunised ID with either gp140 alone or together with Poly(I:C) or Ampligen. On day 28, gp140-specific IgG and IgA were titrated in serum ( a ), vaginal fluids ( b ) and fecal extract ( c ). Data represented mean+s.e.m. of Ab titer from three pooled experiments representing a total of 10–24 mice per group. Statistics using the Kruskal–Wallis test and Dunn’s multiple comparisons.
    Figure Legend Snippet: Ampligen promotes HIV-1 gp140-specific mucosal IgG but not IgA. Mice were immunised ID with either gp140 alone or together with Poly(I:C) or Ampligen. On day 28, gp140-specific IgG and IgA were titrated in serum ( a ), vaginal fluids ( b ) and fecal extract ( c ). Data represented mean+s.e.m. of Ab titer from three pooled experiments representing a total of 10–24 mice per group. Statistics using the Kruskal–Wallis test and Dunn’s multiple comparisons.

    Techniques Used: Mouse Assay

    Adjuvants promoting mucosal Abs to HIV-1 gp140 after ID vaccination. HIV-1 gp140 IgG and IgA were titrated at day 28 in serum ( a ), vaginal fluids ( b ) and fecal extracts ( c ) from mice immunised ID with gp140 alone or in the presence of either Poly(I:C), CpG, DC-Chol or Imiquimod (Aldara). Results are expressed as mean+s.e.m. of Ab titers from pooled experiments representing a total of 12 (DC-chol, Aldara), 20 (CpG) and 80 (gp140 alone, gp140+Poly(I:C)) mice. Statistics using the Kruskall–Wallis test and Dunn’s multiple comparisons.
    Figure Legend Snippet: Adjuvants promoting mucosal Abs to HIV-1 gp140 after ID vaccination. HIV-1 gp140 IgG and IgA were titrated at day 28 in serum ( a ), vaginal fluids ( b ) and fecal extracts ( c ) from mice immunised ID with gp140 alone or in the presence of either Poly(I:C), CpG, DC-Chol or Imiquimod (Aldara). Results are expressed as mean+s.e.m. of Ab titers from pooled experiments representing a total of 12 (DC-chol, Aldara), 20 (CpG) and 80 (gp140 alone, gp140+Poly(I:C)) mice. Statistics using the Kruskall–Wallis test and Dunn’s multiple comparisons.

    Techniques Used: Mouse Assay

    ID vaccination with HSV-2 gD glycoprotein with Poly(I:C) induces mucosal and systemic immunity and protects from lethal vaginal HSV-2 infection. ( a ) Mice were immunised ID with HSV-2 gD alone or together with Poly(I:C) or Ampligen and gD-specific IgG and IgA were measured 7 days after the last immunisation, in serum, vaginal fluids and fecal extracts. Data represent mean+s.e.m. of Ab titres of six experiments each using 6–7 mice per group. ( b – e ) Mice were injected three times ID with either PBS, gD alone, gD+Poly(I:C), gD+Ampligen or once ivag with HSV-2 tk − . One week after the last immunisation, mice were challenged ivag with 10 4 plaque-forming units (PFU) of virulent HSV-2 and protection against infection was followed by body weight change ( b ), survival ( c ), clinical score ( d ) and neurological score ( e ). Data correspond to a pool of two experiments ( b – e ) each with 6–7 mice per group. Statistical analyses were performed using the Kruskall Wallis test and Dunn’s multiple comparisons ( a , e ), the two-way analysis of variance (ANOVA) test and Bonferroni’s multiple comparisons ( b , d ) and the Gehan–Breslow–Wilcoxon test ( c ). Asterisks ( b – d ) indicate significance compared with the PBS control group.
    Figure Legend Snippet: ID vaccination with HSV-2 gD glycoprotein with Poly(I:C) induces mucosal and systemic immunity and protects from lethal vaginal HSV-2 infection. ( a ) Mice were immunised ID with HSV-2 gD alone or together with Poly(I:C) or Ampligen and gD-specific IgG and IgA were measured 7 days after the last immunisation, in serum, vaginal fluids and fecal extracts. Data represent mean+s.e.m. of Ab titres of six experiments each using 6–7 mice per group. ( b – e ) Mice were injected three times ID with either PBS, gD alone, gD+Poly(I:C), gD+Ampligen or once ivag with HSV-2 tk − . One week after the last immunisation, mice were challenged ivag with 10 4 plaque-forming units (PFU) of virulent HSV-2 and protection against infection was followed by body weight change ( b ), survival ( c ), clinical score ( d ) and neurological score ( e ). Data correspond to a pool of two experiments ( b – e ) each with 6–7 mice per group. Statistical analyses were performed using the Kruskall Wallis test and Dunn’s multiple comparisons ( a , e ), the two-way analysis of variance (ANOVA) test and Bonferroni’s multiple comparisons ( b , d ) and the Gehan–Breslow–Wilcoxon test ( c ). Asterisks ( b – d ) indicate significance compared with the PBS control group.

    Techniques Used: Infection, Mouse Assay, Injection

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    Article Snippet: .. Titers of IgG in serum were determined by sandwich ELISA using specific goat anti–mouse polyclonal reagents (Southern Biotechnology Associates, Inc.). .. Purified IgG was used for the standard curve and for calculation of Ab concentration.

    Article Title: Efficient Peripheral Clonal Elimination of B Lymphocytes in MRL/lpr Mice Bearing Autoantibody Transgenes
    Article Snippet: Sera were taken at 6–8 wk of age (young mice) or 5 mo of age (aged mice). .. Total IgG and IgM concentrations were measured by a sandwich ELISA using specific goat anti–mouse polyclonal reagents (Southern Biotechnology Associates, Birmingham, AL). .. Unlabeled goat anti–mouse antibodies (10 μg/ml) in PBS were adhered to polystyrene Immunolon microtiter plates (Dynatech Laboratories, Inc., Chantilly, VA) by overnight incubation.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Selection at Multiple Checkpoints Focuses VH12 B Cell Differentiation toward a Single B-1 Cell Specificity
    Article Snippet: Therefore, we cotransfected into P3-X63-Ag8.653 myeloma cells with the 10/G4 or 2-12 expression constructs with L chain expression vectors containing Vκ4/5H or Vκ21C rearrangements as described . .. To test whether a complete Ig molecule was formed, supernatant was subjected to ELISA using microtiter plates coated with polyclonal goat anti–mouse μ (Southern Biotechnology Associates, Inc.) and alkaline phosphatase–labeled polyclonal goat anti–mouse κ (Southern Biotechnology Associates, Inc.) to develop the reaction. .. In those cases where Ig secretion was not detected, the production of H and L chains was confirmed by ELISA using cell lysates and the polyclonal goat anti–mouse μ– or polyclonal goat anti–mouse κ-coated plates as above.

    Article Title: Selection at Multiple Checkpoints Focuses VH12 B Cell Differentiation toward a Single B-1 Cell Specificity
    Article Snippet: To test whether a complete Ig molecule was formed, supernatant was subjected to ELISA using microtiter plates coated with polyclonal goat anti–mouse μ (Southern Biotechnology Associates, Inc.) and alkaline phosphatase–labeled polyclonal goat anti–mouse κ (Southern Biotechnology Associates, Inc.) to develop the reaction. .. In those cases where Ig secretion was not detected, the production of H and L chains was confirmed by ELISA using cell lysates and the polyclonal goat anti–mouse μ– or polyclonal goat anti–mouse κ-coated plates as above. ..

    Expressing:

    Article Title: CXCR5 CAR-T cells simultaneously target B cell non-Hodgkin’s lymphoma and tumor-supportive follicular T helper cells
    Article Snippet: 2) Monocytes and DCs: FITC anti-CD1c (L161, #331518, 1:100), PE anti-CXCR5 (51505, #FAB190P-100, R & D Systems/Bio-Techne, 1:10), PE/Cy7 anti-CD11c (Bu15, #337216, 1:100), APC anti-CD14 (HCD14, #325608, 1:100), APC/Fire750 anti-CD303 (201A, #354236, 1:100), BV421 anti-HLA-DR (L243, #307636, 1:100), BV510 anti-CD19 (SJ25C1, #363020, 1:100) and BV510 anti-CD3 (SK7, #344828, 1:100). .. CAR expression on transduced mouse splenocytes was detected using polyclonal PE-labeled goat anti-mouse IgG-Ab (#1030-09, Southern Biotech/Biozol, 1:200). ..

    Staining:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

    Modification:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

    Avidin-Biotin Assay:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

    Blocking Assay:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

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    SouthernBiotech alkaline phosphatase conjugated goat anti mouse igg
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    Alkaline Phosphatase Conjugated Goat Anti Mouse Igg, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SouthernBiotech ap conjugated goat anti mouse igm
    Follicular Treg (Tfr) cell differentiation and prevention of autoantibody production depend on STIM1/STIM2. a Volcano plot of DEG in unstimulated WT and Stim1/2 -deficient Treg cells overlaid with upregulated (red) or downregulated (blue) genes of the ‘Tfr cell signature’ 8 . b Analysis of Tfr cells in the spleen and LNs of male WT and Stim1/2 Foxp3 mice; means ± SEM of four mice. c Representative immunofluorescence of PNA + germinal centers (GCs) in submandibular LNs of untreated male WT and Stim1/2 Foxp3 mice; scale bar: 200 µm. d Analysis of GC B cells in male WT and Stim1/2 Foxp3 mice; means ± SEM of four mice. e Detection of IgG autoantibodies in the sera of male WT and Stim1/2 Foxp3 mice using a multiplexed auto-antigen array. Sera of 9–11 individual mice were tested. f Analysis of anti-red blood cell <t>(RBC)</t> IgG and <t>IgM</t> autoantibodies using ELISA; means ± SEM of six mice. g Detection of relative IgG binding to RBCs of male WT and Stim1/2 Foxp3 mice by flow cytometry; means of eight mice. h Representative blood smears of WT and Stim1/2 Foxp3 mice; scale bar: 10 µm. i Analyses of RBC counts and hemoglobin levels in the blood of young (
    Ap Conjugated Goat Anti Mouse Igm, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SouthernBiotech goat anti mouse igg2a conjugates
    Representative histologic appearance and immune deposits in BALB/c mice implanted with transfectoma or hybridoma cells secreting 6-19 WT, T228P hinge mutant, or Tg IgA mAb. Note segmental expansion of mesangial matrix and focal proliferation of mesangial cells in mice implanted with WT 6-19 IgA–secreting cells, but essentially normal histologic appearance of glomeruli in mice implanted with cells secreting 6-19 T228P or Tg-1 IgA (periodic acid–Schiff staining). Immunohistochemical analysis reveals extensive fine granular deposits of IgA, <t>IgG2a,</t> and C3 in the mesangium of mice implanted with WT 6-19 IgA–secreting cells, but only minimal extent of these deposits in mice implanted with cells secreting 6-19 T228P or Tg-1 IgA. Because similar results are obtained with the two other 6-19 Tg IgA mAbs, only results obtained with 6-19 Tg-1 IgA are shown. Original magnification, ×400.
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    SouthernBiotech goat anti rat ig alkaline phosphatase conjugate
    ELISA analysis of the interaction between rCfC1qDC and human heat-aggregated IgG. Plates were coated with various PAMPs several concentrations of rCfC1qDC and rTrx, and then incubated with human heat-aggregated IgG. The interaction was detected with <t>goat–anti-human</t> <t>Ig-alkaline</t> <t>phosphatase</t> <t>conjugate</t> at 405 nm. Results are representative of average three such experiments.
    Goat Anti Rat Ig Alkaline Phosphatase Conjugate, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    DP cDC control the induction of the mucosal Ab response to sFliC. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized (N.I) or primed and boosted with sFliC. The Ab response was evaluated 4 d post-boost. (a) Representative plots and absolute number (graphs) of plasma cells (TCRb - CD19 + B220 low CD138 + ) in the MLN. (b) ELISPOT analysis of sFliC IgG and IgA responses in the MLN and SI-LP. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are mean +SD (n=4 mice/group) and are from 1 representative experiment of three performed. *** p

    Journal: Mucosal immunology

    Article Title: CD103+CD11b+ mucosal classical dendritic cells initiate long-term switched antibody responses to flagellin

    doi: 10.1038/mi.2017.105

    Figure Lengend Snippet: DP cDC control the induction of the mucosal Ab response to sFliC. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized (N.I) or primed and boosted with sFliC. The Ab response was evaluated 4 d post-boost. (a) Representative plots and absolute number (graphs) of plasma cells (TCRb - CD19 + B220 low CD138 + ) in the MLN. (b) ELISPOT analysis of sFliC IgG and IgA responses in the MLN and SI-LP. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are mean +SD (n=4 mice/group) and are from 1 representative experiment of three performed. *** p

    Article Snippet: Following incubation for 1 h at 37°C, plate-bound antibodies were detected using alkaline phosphatase conjugated goat anti-mouse IgG, IgG1, and IgA (Southern Biotech).

    Techniques: Mouse Assay, Enzyme-linked Immunospot

    The mucosal response to sFliC contributes to the systemic antibody response. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized or sFliC prime-boosted. (a) ELISPOT analysis of sFliC IgG and IgA responses in the BM. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are shown as mean +SD (n=12 mice/group) of three independent experiments pooled together. *** p

    Journal: Mucosal immunology

    Article Title: CD103+CD11b+ mucosal classical dendritic cells initiate long-term switched antibody responses to flagellin

    doi: 10.1038/mi.2017.105

    Figure Lengend Snippet: The mucosal response to sFliC contributes to the systemic antibody response. Irf4 fl/fl (black) or Cd11c-cre.Irf4 fl/fl (white) mice were either non-immunized or sFliC prime-boosted. (a) ELISPOT analysis of sFliC IgG and IgA responses in the BM. Number of spot-forming units (SFU) per 5x10 5 cells (graphs) and representative pictures of wells (lower panels). Data are shown as mean +SD (n=12 mice/group) of three independent experiments pooled together. *** p

    Article Snippet: Following incubation for 1 h at 37°C, plate-bound antibodies were detected using alkaline phosphatase conjugated goat anti-mouse IgG, IgG1, and IgA (Southern Biotech).

    Techniques: Mouse Assay, Enzyme-linked Immunospot

    Follicular Treg (Tfr) cell differentiation and prevention of autoantibody production depend on STIM1/STIM2. a Volcano plot of DEG in unstimulated WT and Stim1/2 -deficient Treg cells overlaid with upregulated (red) or downregulated (blue) genes of the ‘Tfr cell signature’ 8 . b Analysis of Tfr cells in the spleen and LNs of male WT and Stim1/2 Foxp3 mice; means ± SEM of four mice. c Representative immunofluorescence of PNA + germinal centers (GCs) in submandibular LNs of untreated male WT and Stim1/2 Foxp3 mice; scale bar: 200 µm. d Analysis of GC B cells in male WT and Stim1/2 Foxp3 mice; means ± SEM of four mice. e Detection of IgG autoantibodies in the sera of male WT and Stim1/2 Foxp3 mice using a multiplexed auto-antigen array. Sera of 9–11 individual mice were tested. f Analysis of anti-red blood cell (RBC) IgG and IgM autoantibodies using ELISA; means ± SEM of six mice. g Detection of relative IgG binding to RBCs of male WT and Stim1/2 Foxp3 mice by flow cytometry; means of eight mice. h Representative blood smears of WT and Stim1/2 Foxp3 mice; scale bar: 10 µm. i Analyses of RBC counts and hemoglobin levels in the blood of young (

    Journal: Nature Communications

    Article Title: Tissue resident and follicular Treg cell differentiation is regulated by CRAC channels

    doi: 10.1038/s41467-019-08959-8

    Figure Lengend Snippet: Follicular Treg (Tfr) cell differentiation and prevention of autoantibody production depend on STIM1/STIM2. a Volcano plot of DEG in unstimulated WT and Stim1/2 -deficient Treg cells overlaid with upregulated (red) or downregulated (blue) genes of the ‘Tfr cell signature’ 8 . b Analysis of Tfr cells in the spleen and LNs of male WT and Stim1/2 Foxp3 mice; means ± SEM of four mice. c Representative immunofluorescence of PNA + germinal centers (GCs) in submandibular LNs of untreated male WT and Stim1/2 Foxp3 mice; scale bar: 200 µm. d Analysis of GC B cells in male WT and Stim1/2 Foxp3 mice; means ± SEM of four mice. e Detection of IgG autoantibodies in the sera of male WT and Stim1/2 Foxp3 mice using a multiplexed auto-antigen array. Sera of 9–11 individual mice were tested. f Analysis of anti-red blood cell (RBC) IgG and IgM autoantibodies using ELISA; means ± SEM of six mice. g Detection of relative IgG binding to RBCs of male WT and Stim1/2 Foxp3 mice by flow cytometry; means of eight mice. h Representative blood smears of WT and Stim1/2 Foxp3 mice; scale bar: 10 µm. i Analyses of RBC counts and hemoglobin levels in the blood of young (

    Article Snippet: After blocking, serial dilutions of mouse sera were incubated, washed, and bound anti-RBC autoantibodies were detected using AP-conjugated goat-anti-mouse IgM and IgG as detection antibodies (Southern Biotech).

    Techniques: Cell Differentiation, Mouse Assay, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Cytometry

    Representative histologic appearance and immune deposits in BALB/c mice implanted with transfectoma or hybridoma cells secreting 6-19 WT, T228P hinge mutant, or Tg IgA mAb. Note segmental expansion of mesangial matrix and focal proliferation of mesangial cells in mice implanted with WT 6-19 IgA–secreting cells, but essentially normal histologic appearance of glomeruli in mice implanted with cells secreting 6-19 T228P or Tg-1 IgA (periodic acid–Schiff staining). Immunohistochemical analysis reveals extensive fine granular deposits of IgA, IgG2a, and C3 in the mesangium of mice implanted with WT 6-19 IgA–secreting cells, but only minimal extent of these deposits in mice implanted with cells secreting 6-19 T228P or Tg-1 IgA. Because similar results are obtained with the two other 6-19 Tg IgA mAbs, only results obtained with 6-19 Tg-1 IgA are shown. Original magnification, ×400.

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: O-Linked Glycosylation Determines the Nephritogenic Potential of IgA Rheumatoid Factor

    doi: 10.1681/ASN.2013070771

    Figure Lengend Snippet: Representative histologic appearance and immune deposits in BALB/c mice implanted with transfectoma or hybridoma cells secreting 6-19 WT, T228P hinge mutant, or Tg IgA mAb. Note segmental expansion of mesangial matrix and focal proliferation of mesangial cells in mice implanted with WT 6-19 IgA–secreting cells, but essentially normal histologic appearance of glomeruli in mice implanted with cells secreting 6-19 T228P or Tg-1 IgA (periodic acid–Schiff staining). Immunohistochemical analysis reveals extensive fine granular deposits of IgA, IgG2a, and C3 in the mesangium of mice implanted with WT 6-19 IgA–secreting cells, but only minimal extent of these deposits in mice implanted with cells secreting 6-19 T228P or Tg-1 IgA. Because similar results are obtained with the two other 6-19 Tg IgA mAbs, only results obtained with 6-19 Tg-1 IgA are shown. Original magnification, ×400.

    Article Snippet: The precipitates were solubilized in PBS containing 1% BSA and 0.05% Tween-20 and subjected to ELISA using plates coated with goat anti-mouse IgA (Bethy Laboratories, Inc., Cambridge, UK) and the assay was developed with alkaline phosphatase–labeled goat anti-mouse IgG2a conjugates (Southern Biotechnology Associates, Inc., Birmingham, AL).

    Techniques: Mouse Assay, Mutagenesis, Staining, Immunohistochemistry

    ELISA analysis of the interaction between rCfC1qDC and human heat-aggregated IgG. Plates were coated with various PAMPs several concentrations of rCfC1qDC and rTrx, and then incubated with human heat-aggregated IgG. The interaction was detected with goat–anti-human Ig-alkaline phosphatase conjugate at 405 nm. Results are representative of average three such experiments.

    Journal: PLoS ONE

    Article Title: A C1q Domain Containing Protein from Scallop Chlamys farreri Serving as Pattern Recognition Receptor with Heat-Aggregated IgG Binding Activity

    doi: 10.1371/journal.pone.0043289

    Figure Lengend Snippet: ELISA analysis of the interaction between rCfC1qDC and human heat-aggregated IgG. Plates were coated with various PAMPs several concentrations of rCfC1qDC and rTrx, and then incubated with human heat-aggregated IgG. The interaction was detected with goat–anti-human Ig-alkaline phosphatase conjugate at 405 nm. Results are representative of average three such experiments.

    Article Snippet: After SDS-PAGE, the samples of whole cell lysate (4 h after IPTG added) were electrophoretically transferred on to a 0.45 mm pore nitrocellulose membrane at 200 mA for 5 h. The membrane was blocked with PBS containing 3% BSA at 37°C for 1 h, and incubated with antibody (diluted 1∶1000 in PBS) at 37°C for 1 h. After washed three times with PBS containing 0.05% Tween-20 (PBS-T), the membrane was then incubated with goat-anti-rat Ig-alkaline phosphatase conjugate (Southern Biotech) diluted 1∶4000 in PBS at 37°C for 1 h, and then washed three times again with PBS-T.

    Techniques: Enzyme-linked Immunosorbent Assay, Incubation

    ELISA analysis of the interaction between rCfC1qDC and the PAMPs. Plates were coated with various PAMPs, and then incubated with several concentrations of rCfC1qDC and rTrx at 18°C for 3 h. After incubated with rat polyclonal antiserum, the interaction was detected with goat–anti-rat Ig-alkaline phosphatase conjugate at 405 nm. Samples with P/N > 2.1 were considered positive. Results are representative of average three such experiments.

    Journal: PLoS ONE

    Article Title: A C1q Domain Containing Protein from Scallop Chlamys farreri Serving as Pattern Recognition Receptor with Heat-Aggregated IgG Binding Activity

    doi: 10.1371/journal.pone.0043289

    Figure Lengend Snippet: ELISA analysis of the interaction between rCfC1qDC and the PAMPs. Plates were coated with various PAMPs, and then incubated with several concentrations of rCfC1qDC and rTrx at 18°C for 3 h. After incubated with rat polyclonal antiserum, the interaction was detected with goat–anti-rat Ig-alkaline phosphatase conjugate at 405 nm. Samples with P/N > 2.1 were considered positive. Results are representative of average three such experiments.

    Article Snippet: After SDS-PAGE, the samples of whole cell lysate (4 h after IPTG added) were electrophoretically transferred on to a 0.45 mm pore nitrocellulose membrane at 200 mA for 5 h. The membrane was blocked with PBS containing 3% BSA at 37°C for 1 h, and incubated with antibody (diluted 1∶1000 in PBS) at 37°C for 1 h. After washed three times with PBS containing 0.05% Tween-20 (PBS-T), the membrane was then incubated with goat-anti-rat Ig-alkaline phosphatase conjugate (Southern Biotech) diluted 1∶4000 in PBS at 37°C for 1 h, and then washed three times again with PBS-T.

    Techniques: Enzyme-linked Immunosorbent Assay, Incubation