mouse anti pig igg2  (Bio-Rad)

Journal: Frontiers in Immunology

Article Title: Larval ascariasis elicits a prominent IgA and IgG1/2 antibody response to adult Ascaris excretory/secretory antigens in pigs

doi: 10.3389/fimmu.2025.1606128

Figure Lengend Snippet: Antibody isotype responses against A . suum L3, adult and adult ES antigens in serum and bile 35 dpi. (A) Schematic illustrating experimental infection of the pigs. Eight pigs were infected with a single dose of 4000 infective A . suum eggs. In parallel, four uninfected controls were included. Box plots illustrating median serum (B) IgM, IgG, IgA and (C) IgG1 and IgG2 subclass responses against A . suum L3 lysate, adult lysate and adult ES products. (D) Box plots illustrating median bile IgM, IgG and IgA responses against A . suum L3 lysate, adult lysate and adult ES products. The data is reported in arbitrary ELISA units (AEU). Whiskers indicate 95% percentile. Significance assessed using the Wilcoxon test and depicted by p<0.05: *p<0.01: **p<0.001: ***p<0.0001: ****. Ascaris -infected n= 16 and uninfected controls n= 8.

Article Snippet: For IgM, IgA, IgG1 and IgG2, non-conjugated mouse anti-pig IgM (Bio-Rad, # MCA637GA) diluted at 1:20000, mouse anti-pig IgA (Bio-Rad, # MCA638GA) diluted at 1:1000, mouse anti-pig IgG1 (Bio-Rad, # MCA635GA) diluted at 1:1000 and mouse anti-pig IgG2 (Bio-Rad, # MCA638GA) diluted at 1:1000, mouse anti-pig sIgA (Bio-Rad, # MCA634GA) diluted at 1:10000 was used.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Larval ascariasis elicits a prominent IgA and IgG1/2 antibody response to adult Ascaris excretory/secretory antigens in pigs

doi: 10.3389/fimmu.2025.1606128

Figure Lengend Snippet: Comparable Ascaris -specific antibody responses in Ascaris single- and co-infected pigs. (A) Schematic illustrating experimental infection of the pigs. Box plots illustrating median serum IgM, IgG, IgA, IgG1 and IgG2 responses (B) and median bile IgM, IgG and IgA responses (C) against A . suum adult ES products. (D) Box plots illustrating L3- and adult-specific IgA responses in bile and serum. Blue, red, orange and green represent uninfected controls, A. suum infected, A . suum- S . Typhimurium coinfected and S. Typhimurium infected pigs respectively. Whiskers indicate 95% percentile. Significance is represented by p<0.05: *p<0.01: **p<0.001: ***p<0.0001: ****. Significance was tested using Kruskal Wallis with Dunn or Games-Howell test and Wilcoxon test. Uninfected controls n = 12, Ascaris single infected n = 12, Ascaris-Salmonella coinfected n = 12, Salmonella single infected n =12.

Article Snippet: For IgM, IgA, IgG1 and IgG2, non-conjugated mouse anti-pig IgM (Bio-Rad, # MCA637GA) diluted at 1:20000, mouse anti-pig IgA (Bio-Rad, # MCA638GA) diluted at 1:1000, mouse anti-pig IgG1 (Bio-Rad, # MCA635GA) diluted at 1:1000 and mouse anti-pig IgG2 (Bio-Rad, # MCA638GA) diluted at 1:1000, mouse anti-pig sIgA (Bio-Rad, # MCA634GA) diluted at 1:10000 was used.

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Larval ascariasis elicits a prominent IgA and IgG1/2 antibody response to adult Ascaris excretory/secretory antigens in pigs

doi: 10.3389/fimmu.2025.1606128

Figure Lengend Snippet: Strong positive correlation between sIgA against A . suum adult ES and eosinophil influx. (A) Pearson correlation of sIgA against adult ES products and %eosinophil frequencies of leukocytes in BAL . (B) Box plots illustrating median IgM, IgG and IgA responses against A . suum adult ES products in BAL fluid. (C) Box plots illustrating sIgA against L3 lysate, adult lysate and adult ES in BAL fluid. Blue and red denotes uninfected controls and A. suum infected pigs, respectively. Whiskers indicate 95% percentile. Significance determined by Wilcoxon test is represented by p<0.05: *p<0.01: **.

Article Snippet: For IgM, IgA, IgG1 and IgG2, non-conjugated mouse anti-pig IgM (Bio-Rad, # MCA637GA) diluted at 1:20000, mouse anti-pig IgA (Bio-Rad, # MCA638GA) diluted at 1:1000, mouse anti-pig IgG1 (Bio-Rad, # MCA635GA) diluted at 1:1000 and mouse anti-pig IgG2 (Bio-Rad, # MCA638GA) diluted at 1:1000, mouse anti-pig sIgA (Bio-Rad, # MCA634GA) diluted at 1:10000 was used.

Techniques: Infection

Journal: Antiviral research

Article Title: Targeted mutagenesis of the β-strand DNA binding region of African swine fever virus histone-like protein (pA104R) impairs DNA-binding activity and antibody recognition.

doi: 10.1016/j.antiviral.2023.105784

Figure Lengend Snippet: Fig. 4. Mapping of pA104R B-cell epitopes. Pooled porcine sera (n = 82), at optimized dilutions of 1:250 (IgM) and 1:500 (IgG), were subjected to peptide-based ELISA. (A) Three IgM immunodominant regions were identified producing a clear increase in antibody titers on day 7 p.i. that remains high at least until day 16 p.i. (B) Two IgG immunodominant regions were likewise identified that produce a clear increase in antibody titers on day 13 p.i. that remains high at least until day 35 p. i. Data are presented as mean ± SD of 3 replicates. The cut-off value (CO) was calculated as mean OD of the negative controls (n = 11). If the mean OD of negative controls was lower than 0.105, the value was considered to be 0.105. Values above the solid black lines (S/CO > 1 IgM and S/CO > 2 IgG) were scored as weakly positive and values above the dotted lines (S/CO > 2 IgM and S/CO > 4 IgG) were scored as strongly positive reactions. Grey bars represent weakly positive porcine anti-pA104R epitopes and black bars represent strongly positive porcine anti-PA104R epitopes. DPI – days post-infection.

Article Snippet: The plates were incubated with 1:200 diluted pig sera in 5% BSA in PBS for 1 h, followed by a blocking step with 5% BSA diluted in PBS for 1 h. The plates were then incubated with the 1:100 diluted polyclonal mouse anti-pig IgG1 antibody clone K139 3C8 (BioRad) or polyclonal mouse anti-pig IgG2 antibody clone K68 Ig2 (1:1000) (Bio-Rad) primary antibodies for 1 h, followed by incubation with the HRP-conjugated goat anti-mouse secondary antibody (1:5000) (BioRad) for an additional hour.

Techniques: Peptide ELISA, Infection

Journal: Antiviral research

Article Title: Targeted mutagenesis of the β-strand DNA binding region of African swine fever virus histone-like protein (pA104R) impairs DNA-binding activity and antibody recognition.

doi: 10.1016/j.antiviral.2023.105784

Figure Lengend Snippet: Fig. 5. Analysis of anti-pA104R antibodies recognizing linear B-cell epitopes. (A) The IgM antibody determinants identified from infected porcine sera at 7, 10, and 13–16 days post-infection. (B) The IgG antibody determinants identified from infected porcine sera at 13–16, 17–20, and 21–30 days post-infection. Regions of amino acid sequences corresponding to the identified B-cell epitopes are indicated in the schematic diagrams of the pA104R sequence. The percentage of antibody recognition contributed by each individual pA104R epitope is indicated in the pie charts and was calculated according to the following equation: % antibody recognition = 100 × (OD values from individual peptide/sum of the OD values from all peptides). In this calculation, the avidity and affinity of the peptides to the sera were assumed to be similar.

Article Snippet: The plates were incubated with 1:200 diluted pig sera in 5% BSA in PBS for 1 h, followed by a blocking step with 5% BSA diluted in PBS for 1 h. The plates were then incubated with the 1:100 diluted polyclonal mouse anti-pig IgG1 antibody clone K139 3C8 (BioRad) or polyclonal mouse anti-pig IgG2 antibody clone K68 Ig2 (1:1000) (Bio-Rad) primary antibodies for 1 h, followed by incubation with the HRP-conjugated goat anti-mouse secondary antibody (1:5000) (BioRad) for an additional hour.

Techniques: Infection, Sequencing

Journal: Antiviral research

Article Title: Targeted mutagenesis of the β-strand DNA binding region of African swine fever virus histone-like protein (pA104R) impairs DNA-binding activity and antibody recognition.

doi: 10.1016/j.antiviral.2023.105784

Figure Lengend Snippet: Fig. 7. pA104R-specific IgG reactivity (A) pA104R-specific IgG antibody detection. (B) PEP15-specific IgG antibody detection. (C) PEP15-specific IgG1 and IgG2 antibody isotype detection. (D) IgG1/IgG2 ratio calculated for each IgG sample as the OD value of IgG1 divided by the OD value of IgG2. Red solid lines represent medians. DPI = days post-infection.

Article Snippet: The plates were incubated with 1:200 diluted pig sera in 5% BSA in PBS for 1 h, followed by a blocking step with 5% BSA diluted in PBS for 1 h. The plates were then incubated with the 1:100 diluted polyclonal mouse anti-pig IgG1 antibody clone K139 3C8 (BioRad) or polyclonal mouse anti-pig IgG2 antibody clone K68 Ig2 (1:1000) (Bio-Rad) primary antibodies for 1 h, followed by incubation with the HRP-conjugated goat anti-mouse secondary antibody (1:5000) (BioRad) for an additional hour.

Techniques: Infection

Journal: Antiviral research

Article Title: Targeted mutagenesis of the β-strand DNA binding region of African swine fever virus histone-like protein (pA104R) impairs DNA-binding activity and antibody recognition.

doi: 10.1016/j.antiviral.2023.105784

Figure Lengend Snippet: Fig. 6. PEP-specific IgM and IgG antibody determinants. (A) PEP-specific IgM antibody detection in sera (n = 45) at a dilution of 1:250 was determined by peptide- based ELISA. Sera were categorized into non-infected or days post-infection (DPI) = 0 (n = 10), DPI = 7 (n = 10), DPI = 10 (n = 9), and DPI = 13–16 (n = 16). Red solid lines represent medians. (B) PEP-specific IgG antibody detection in sera (n = 48) at a dilution of 1:500 was determined by peptide-based ELISA. Sera were categorized into non-infected or DPI = 0 (n = 10), DPI = 13–16 (n = 16), DPI = 17–20 (n = 11), and DPI = 21–30 (n = 11). Red solid lines represent medians. (C) The pA104R dimeric structure (PDB 6LMH), generated using UCSF Chimera software. The IgM and IgG antibody determinants are indicated in brown (PEP23), and blue (PEP15).

Article Snippet: The plates were incubated with 1:200 diluted pig sera in 5% BSA in PBS for 1 h, followed by a blocking step with 5% BSA diluted in PBS for 1 h. The plates were then incubated with the 1:100 diluted polyclonal mouse anti-pig IgG1 antibody clone K139 3C8 (BioRad) or polyclonal mouse anti-pig IgG2 antibody clone K68 Ig2 (1:1000) (Bio-Rad) primary antibodies for 1 h, followed by incubation with the HRP-conjugated goat anti-mouse secondary antibody (1:5000) (BioRad) for an additional hour.

Techniques: Peptide ELISA, Infection, Generated, Software