biotinylated-anti-il21 antibody  (R&D Systems)

Journal: PLoS ONE

Article Title: Interleukin 6 Accelerates Mortality by Promoting the Progression of the Systemic Lupus Erythematosus-Like Disease of BXSB. Yaa Mice

doi: 10.1371/journal.pone.0153059

Figure Lengend Snippet: Sera from BXSB. Yaa and BXSB mice of different ages were collected and tested for the levels of (A) IL6 (B) anti-nuclear antibodies (ANA) and anti-dsDNA antibodies, and (C) immunoglobulin isotypes (IgG2b and IgG2c); n = 4–20 mice per age group. Each dot represents one mouse. Horizontal bars represent mean ±SEM. p values are calculated by one-way ANOVA with Tukey’s multiple comparison tests. * p<0.05, ** p<0.01, ***p<0.0001.

Article Snippet: IL6 or IL21 were captured by secondary biotinylated anti-mouse IL6 antibody (BD Biosciences) or anti-mouse IL21 antibody (Peprotech) followed by streptavidin-HRP/TMB (Sigma/Invitrogen) for colorimetric estimation.

Techniques: Comparison

Journal: PLoS ONE

Article Title: Interleukin 6 Accelerates Mortality by Promoting the Progression of the Systemic Lupus Erythematosus-Like Disease of BXSB. Yaa Mice

doi: 10.1371/journal.pone.0153059

Figure Lengend Snippet: (A) CD43 - B cells were purified from spleen, BM and peritoneal lavage of 4–5 month old BXSB. Yaa and BXSB mice and stimulated with graded concentrations of Imiquimod (R837). (B) Dendritic cells enriched from spleen (splenic DCs) or differentiated from BM precursors (GMCSF (40ng/ml) and IL4 (20ng/ml) for 9 d with media changed every third day), peritoneal macrophages and CD115 + monocytes from spleens of BXSB. Yaa and BXSB mice were stimulated with R837 (50ng/ml) in vitro . Supernatants were collected after 24 hours and IL6 levels were quantified using a standard sandwich ELISA. Data expressed as mean ± SEM of triplicate wells are representative of 4–6 independent experiments. p values are determined by two-way ANOVA. * p<0.05, ** p<0.01, ***p<0.0001.

Article Snippet: IL6 or IL21 were captured by secondary biotinylated anti-mouse IL6 antibody (BD Biosciences) or anti-mouse IL21 antibody (Peprotech) followed by streptavidin-HRP/TMB (Sigma/Invitrogen) for colorimetric estimation.

Techniques: Purification, In Vitro, Sandwich ELISA

Journal: PLoS ONE

Article Title: Interleukin 6 Accelerates Mortality by Promoting the Progression of the Systemic Lupus Erythematosus-Like Disease of BXSB. Yaa Mice

doi: 10.1371/journal.pone.0153059

Figure Lengend Snippet: (A) Purified CD43 - B cells from 4–5 month old BXSB. Yaa and BXSB mice were stimulated with R837 (50ng/ml) for 24 hours and stained for surface expression of the activation markers—CD40, MHCII, ICOSL and CD86. Data are expressed as mean ± SEM of mean fluorescence intensity (MFI) and are pooled from 4–5 independent experiments. (B) Purified CD43 - B cells from 4–5 month old BXSB. Yaa and BXSB mice were stimulated with graded doses of R837 for 24 hours. 3 H thymidine (0.5μCi/well) was added and its incorporation was determined after 16 hours by liquid scintillation counting. Proliferation was determined as counts per minute (CPM). (C-D) CD43 - B cells were stimulated with R837 (50ng/ml) in the presence and absence of anti-BCR antibody. IL6 secretion (C) and B cell proliferation (D) were determined as described earlier. (E) Purified B cells from 4–5 month old BXSB. Yaa and BXSB mice were cultured in the presence or absence of R837 (50ng/ml); IFNα and IFNβ (40U/ml), either alone or in combination, for 24h. Supernatants were collected and quantified for IL6 levels by standard sandwich ELISA. All data (B-E) are expressed as mean ± SEM of triplicate wells and are representative of 2–3 independent experiments. P values are determined by two-way ANOVA. * p<0.05, ** p<0.01, ***p<0.0001.

Article Snippet: IL6 or IL21 were captured by secondary biotinylated anti-mouse IL6 antibody (BD Biosciences) or anti-mouse IL21 antibody (Peprotech) followed by streptavidin-HRP/TMB (Sigma/Invitrogen) for colorimetric estimation.

Techniques: Purification, Staining, Expressing, Activation Assay, Fluorescence, Cell Culture, Sandwich ELISA

Journal: PLoS ONE

Article Title: Interleukin 6 Accelerates Mortality by Promoting the Progression of the Systemic Lupus Erythematosus-Like Disease of BXSB. Yaa Mice

doi: 10.1371/journal.pone.0153059

Figure Lengend Snippet: (A) Sera from 6-month old BXSB. Yaa . Il6 -/- , BXSB. Yaa , BXSB. Il6 -/- , BXSB, BXSB. B6Y and C57BL/6 mice were quantified for the presence of ANA by ELISA (n = 3–10 mice per group). Each dot represents one mouse. Horizontal bars indicate mean ± SEM and p values are calculated by one-way ANOVA with Tukey’s multiple comparison tests. (B) Survival curve analysis of BXSB. Yaa . Il6 +/- (n = 17), BXSB. Yaa . Il6 -/— (n = 41) and archival data of BXSB. Yaa (n = 93) mice. (C-E) IL6-competent and—deficient BXSB. Yaa mice (n = 8–10) were compared for disease manifestations including spleen weight (C), serum IgG1 and IgG2b (D) and serum anti-dsDNA antibody titers (E). Bars represent mean ± SEM of observations from individual mice and p values are calculated using two-way ANOVA. * p<0.05, ** p<0.01, ***p<0.0001.

Article Snippet: IL6 or IL21 were captured by secondary biotinylated anti-mouse IL6 antibody (BD Biosciences) or anti-mouse IL21 antibody (Peprotech) followed by streptavidin-HRP/TMB (Sigma/Invitrogen) for colorimetric estimation.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: PLoS ONE

Article Title: Interleukin 6 Accelerates Mortality by Promoting the Progression of the Systemic Lupus Erythematosus-Like Disease of BXSB. Yaa Mice

doi: 10.1371/journal.pone.0153059

Figure Lengend Snippet: 6–7.5 weeks old BXSB. Yaa (n = 5) and BXSB. Yaa . Il6 -/- (n = 8) mice were bled and (A) cells were stained with fluorescent-labeled antibodies to B220, IgM, MHCII, IL21R and FAS and analyzed by FACS. Expression of MHCII, IL21R and FAS on viable single cells gated on B220 + IgM + is determined as MFI. (B) The frequencies of splenic total B cells (B220 + IgM + ) and GC B cells (B220 + IgM + PNA hi FAS + GL7 + ) were determined and expressed as percentages of total cells. Expression levels of FAS on GC B cells are depicted as mean fluorescence intensity (MFI). Data are expressed as mean ± SEM of individual values obtained from each mouse and statistical analysis is done using two tailed unpaired t test. * p<0.05, ** p<0.01, ***p<0.0001.

Article Snippet: IL6 or IL21 were captured by secondary biotinylated anti-mouse IL6 antibody (BD Biosciences) or anti-mouse IL21 antibody (Peprotech) followed by streptavidin-HRP/TMB (Sigma/Invitrogen) for colorimetric estimation.

Techniques: Staining, Labeling, Expressing, Fluorescence, Two Tailed Test

Journal: PLoS ONE

Article Title: Interleukin 6 Accelerates Mortality by Promoting the Progression of the Systemic Lupus Erythematosus-Like Disease of BXSB. Yaa Mice

doi: 10.1371/journal.pone.0153059

Figure Lengend Snippet: (A) Splenocytes from BXSB. Yaa . Il6 -/- (n = 8) BXSB. Yaa (n = 5), BXSB.B6Y (n = 6) and C57BL/6J (n = 3) mice aged 6–9 weeks were stained with labeled antibodies to CD4, ICOS and PD1. Percentages of CD4 + T cells (CD4 + ) and T FH cells (CD4 + ICOS hi PD1 + ) cells were determined by FACS and expressed as mean ± SEM. (B) Sera from BXSB. Yaa . Il6 -/- (n = 12; 1.5–5 months old), BXSB. Il6 -/- (n = 5; 3–5 months old); BXSB. Yaa (n = 5; 3–5 months old) and BXSB (n = 5; 3–5 months old) mice were quantified for IL21 by sandwich ELISA. Data is expressed as mean ± SEM from observations from each mouse. Statistical analysis was done using one-way ANOVA with Tukey’s multiple comparison test. * p<0.05, ** p<0.01, ***p<0.0001.

Article Snippet: IL6 or IL21 were captured by secondary biotinylated anti-mouse IL6 antibody (BD Biosciences) or anti-mouse IL21 antibody (Peprotech) followed by streptavidin-HRP/TMB (Sigma/Invitrogen) for colorimetric estimation.

Techniques: Staining, Labeling, Sandwich ELISA, Comparison