Ninety-six-well plates were coated with 50 ng per well of recombinant F protein (Sino Biological Inc.) to capture anti-F antibody, or with rabbit anti-mouse IgG (VETOR Laboratories) to capture total mouse IgG, or with 5 Â 10 4 PFU per well of heatinactivated RSV (HI-RSV) to capture RSV-specific IgG in serum. Serially diluted serum were added to the wells and incubated for 2 h. After washing with PBST, HRP-conjugated donkey antimouse IgG antibody (Jackson ImmunoResearch) was added to the wells for 45 min. For detecting anti-RSV IgG, IgG1, and IgG2a, serially diluted samples were added to the plates coated with HI-RSV and then detected by the addition of HRPconjugated goat anti-mouse IgG, anti-mouse IgG1 (Zymed), and anti-mouse IgG2a (Zymed), respectively. The reaction was developed by incubation with 3,3 0 ,5,5 0 -tetramethylbenzidine (TMB) substrate for 20 min in the dark and terminated by adding 2 N H 2 SO 4 . The optical density at 450 nm was determined using a microplate reader.
Anti mouse igg2a
Manufactured by Zymo Research
Anti-mouse IgG2a is a laboratory reagent used to detect and quantify the presence of the IgG2a antibody isotype in mouse samples. It is a highly specific and sensitive antibody that binds to the IgG2a subclass of mouse immunoglobulins.
Automatically generated - may contain errors
2 protocols using anti mouse igg2a
1
ELISA for Antibody Detection
2
Antibody and Cytokine Responses in T. crassiceps Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Blood was obtained by a small cut in the tail vein and centrifuged to obtain serum. Serum was stored at −70 °C until use. Then, 96-well plates (Nunc, Polysorp) were incubated with 10μg/mL of Taenia crassiceps soluble antigen overnight at 4 °C. After blockade, serum from the different groups was adjusted at 1:25 dilution with subsequent double dilutions in PBS. After overnight incubation, anti-mouse IgG1 or anti-mouse IgG2a coupled with HRP (Zymed) was added to the plate. IgG levels were evidenced by adding ABTS/H2O2. For cytokines, sera from STAT1+/+ and STAT1−/− T. crassiceps-infected or naïve mice were obtained and ELISA for IL-13, IL-10, TNF-α, and IL-17 was performed according to the manufacturer’s instructions (Peprotech, Mexico).
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