Feces were collected from Sox8+/+ and Sox8−/− mice. Freeze-dried feces were homogenized in PBS with EDTA-free protease inhibitor cocktail (Complete EDTA-free; Roche) and centrifuged at 4°C. Fecal supernatants were diluted in 2% BSA/PBS. MaxiSorp plates (Thermo Fisher Scientific) were coated with goat anti-mouse IgA (Bethyl Laboratories) for 1 h. The plates were washed five times with 0.1% Tween20 in Tris-buffered saline (TBS-T) and then blocked with 2% BSA/PBS at room temperature for 30 min. The plate was incubated with the diluted samples at room temperature for 1 h. After washing five times with TBS-T, the plate was incubated with HRP-conjugated goat anti-mouse IgA antibodies (Bethyl Laboratories) at room temperature for 1 h. After washing five times with TBS-T, the plate was incubated with 1-Step Ultra TMB-ELISA Substrate Solution (Thermo Fisher Scientific). The reaction was stopped by adding 1.2 M sulfuric acid. The absorbance was measured at 450 nm.
Hrp conjugated goat anti mouse iga antibodies
Manufactured by Fortis Life Sciences
HRP-conjugated goat anti-mouse IgA antibodies are a laboratory reagent used for the detection and quantification of mouse IgA antibodies in various immunoassays. They consist of goat-derived polyclonal antibodies specific to mouse IgA that are conjugated to the enzyme horseradish peroxidase (HRP).
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Lab products found in correlation
1 step ultra tmb elisa substrate solution, by Thermo Fisher Scientific (1 mentions)
Goat anti mouse iga, by Fortis Life Sciences (1 mentions)
Maxisorp plate, by Thermo Fisher Scientific (1 mentions)
Complete edta free, by Roche (1 mentions)
Elisa plate, by Corning (1 mentions)
Hrp conjugated goat anti mouse igg antibody, by Fortis Life Sciences (1 mentions)
Tmb substrate, by BioLegend (1 mentions)
2 protocols using hrp conjugated goat anti mouse iga antibodies
1
Quantifying IgA in Mouse Feces
2
Antibody Quantification in Mouse Sera and BALF
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ELISAs were used to measure antibodies in immunized mouse sera and BALF samples. After coating with 2 μg/mL recombinant HA protein, 96-well ELISA plates (Costar) were incubated overnight at 4 °C, blocked with 1% BSA in PBS at room temperature, and incubated for 30 min. Serially diluted samples were added to each plate and incubated for 1 h at room temperature. Next, HRP-conjugated goat anti-mouse IgG antibodies (1:30,000) or HRP-conjugated goat anti-mouse IgA antibodies (1:10,000) (Bethyl) were added to individual plates and incubated for another 1 h at room temperature. TMB substrates (BioLegend) were added for coloration, followed by incubation for 15 to 20 min at room temperature; reactions were stopped with 2 N H2SO4 and detected using an ELISA reader (OD450). End-point titers were measured as fourfold absorbance of a negative control.
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