Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BM5 by intramuscular (IM) N = 7 and Gene Gun (GG), N = 8 immunization. Antigen-specific ELISAs were performed with sera collected 8 wk after the first immunization. A : IgG1 and IgG2a results shown are mean OD + SE for IM and GG mice; B : IgG1 to IgG2a ratios are shown for mice immunized with BM5 by IM and GG routes. Values shown are mean PD +SE. The ratios in the two groups were significantly different (p < 0.05).

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BMIF by IM (N = 12) and GG (N = 9). Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG1 and IgG2a antibodies shown are mean OD + SE; B: IgG1 to IgG2a ratios are shown for mice immunized with BMIF by IM and GG routes. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BMHSP by IM (N = 12) and GG (N = 9) route. Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG1 and IgG2a results shown are mean OD +SE; B: IgG1 to IgG2a ratios are shown for mice immunized with BMHSP by IM and GG route. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BM14 by IM (N = 12) and GG (N = 9). Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG, IgG1 and IgG2a antibody results shown are mean OD + SE; B: IgG1 to IgG2a antibody ratios are shown for mice immunized with BMHSP by IM and GG routes. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: Time course of total IgG antibodies to individual recombinant B. malayi antigens after polyvalent and monovalent DNA vaccination by IM injection. ELISAs were performed with sera collected at different times after immunization. Results shown are mean OD + SE. A: IgG antibody responses to individual antigens after polyvalent DNA vaccination (5 mice). B: IgG antibody responses to individual antigens after monovalent DNA vaccination (7 mice for BM5, 12 mice for each of the other antigens).

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques: Recombinant, Injection

Journal: Cell reports

Article Title: Complement C4A Regulates Autoreactive B Cells in Murine Lupus

doi: 10.1016/j.celrep.2020.108330

Figure Lengend Snippet: (A) Splenic GC B cell population (B220 + CD38 dim/− GL7 high ) analyzed by flow cytometry in 15 wo C4A, C4B, and C4 +/+ 564Igi. Means ± SEMs; 1-way ANOVA with Tukey’s test; 1 dot represents 1 mouse; n > 6. (B–D) Analysis of GC structures by immunohistochemistry. (B) Representative images of spleen follicles. Marginal zone macrophages: CD169 (red), B cell zone: B220 (blue), T cell zone: CD3 (green) and GC: GL7 (white); the scale bar represents 100 μm. (C) Frequency of GL7 + follicles. Means ± SEMs; 1-way ANOVA with Tukey’s test; 1 dot represents 1 mouse; n > 8 for each 564Igi strain. (D) Quantification of individual GL7 + area using ImageJ software. Means ± SEMs; 1-way ANOVA with Tukey’s test; 1 dot represents 1 GL7 + area; n > 7 animals for each 564Igi strain. (E) Serum anti-nucleoli IgG titers measured by ELISA. Means ± SEMs; 1 dot represents 1 mouse; n > 8. *p < 0.05, **p < 0.01, ****p < 0.0001). See also .

Article Snippet: Polyclonal Goat anti-Mouse IgG2a, Human-absorbed-AP , Southern Biotech , Cat# 1080-04, RRID:AB_2692322.

Techniques: Flow Cytometry, Immunohistochemistry, Software, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: Complement C4A Regulates Autoreactive B Cells in Murine Lupus

doi: 10.1016/j.celrep.2020.108330

Figure Lengend Snippet: (A) Volcano plot comparison of median IgG signal intensities in C4A 564Igi (n = 8) relative to C4B 564Igi (n = 10) mice. Log2(fold change) appears on the x axis. Significance (presented as −10 × log10[p value]) is on the y axis. The p value was obtained running multiple unpaired t tests corrected with the Holm-Sidak method. One dot represents 1 self-Ag. (B–D) Dosage of anti-Ro60, -BPI, and -CTGF IgG titers in serum of 15–20 wo C4 −/− , C4A, C4B, and C4 +/+ 564Igi by ELISA. Means ± SEMs; 1 dot represents 1 mouse; n > 4; 1-way ANOVA with Tukey’s test; *p < 0.05. See also and .

Article Snippet: Polyclonal Goat anti-Mouse IgG2a, Human-absorbed-AP , Southern Biotech , Cat# 1080-04, RRID:AB_2692322.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: Complement C4A Regulates Autoreactive B Cells in Murine Lupus

doi: 10.1016/j.celrep.2020.108330

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Polyclonal Goat anti-Mouse IgG2a, Human-absorbed-AP , Southern Biotech , Cat# 1080-04, RRID:AB_2692322.

Techniques: Purification, Recombinant, Software

Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 6. In vitro antigenicity analysis of t-M2e-t NCs using ELISA. (A) Absorbance values from an indirect ELISA performed by coating 96-well plates with a fixed amount of either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a negative control peptide (NCP). Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of M2e detection IgG antibody, which was applied at different dilutions. (B) Absorbance values from a sandwich ELISA performed by coating 96-well plates with a fixed amount of purified mouse anti-M2e IgM monoclonal antibody as capture antibody. Dilutions of analyte (either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a NCP) were then added to the wells. Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of primary M2e detection IgG antibody. Cartoons of each ELISA procedure are shown below their respective graphs.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Indirect ELISA, Negative Control, Formulation, Sandwich ELISA

Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 7. Immune response and survival data of mice after immunization with t-M2e-t NCs. Mice were immunized thrice, once each on day 0, 21, and 42. Blood was collected and analyzed for anti-M2e antibodies. Within 1 week of the final serum collection, mice were challenged with 3× LD50 A/California/07/2009 (H1N1) and monitored for 14 days. Two antigens were used (i) 20 μg or 5 μg t-M2e-t UCP (un-cross-linked peptide) with/without 20 μg CpG, and (ii) 20 μg or 5 μg S t-M2e-t NCs (small NCs) with/without 20 μg CpG. (A−C) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t UCP as antigen. (D−F) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t NC as antigen. (G, H) Body weight and survival after virus challenge of t-M2e-t UCP groups. (I, J) Body weight and survival after virus challenge of t-M2e-t NP groups. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: Virus