Fitc conjugated anti mouse iga antibody

CH12F3 cells were cultured in RPMI 1640 medium supplemented with 10% (vol/vol) fetal bovine serum and 50 μM β-mercaptoethanol. For CSR assays, cells (viability >95%) were seeded at 5×10⁴ cells/mL in medium containing 1 μg/mL anti-CD40 antibody (eBioscience), 5 ng/mL of IL-4 (R&D Systems), and 0.5 ng/mL TGF-β1 (R&D Systems) and grown for 72 hours. Cells were stained with a FITC-conjugated anti-mouse IgA antibody (BD Biosciences) and analyzed on a LSR II flow cytometer (BD Biosciences). CSR efficiency is determined as the percentage of IgA-positive cells. Rat anti-human/mouse AID antibody used for western blotting was purchased from eBioscience. Anti-AID antibody for chromatin-immunoprecipitation (ChIP) analyses was purchased from Active Motif. ChIP assays were performed as previously described (14 (link)).

CH12F3.2a and its derivative cells were cultured in RPMI medium supplemented with 10% FCS and 50 μM β-mercaptoethanol. As for CSR assay, healthy cells in log phase were seeded at 5 × 10⁴ cells/ml in medium with 1 μg/ml anti-CD40 (eBioscience #16-0404-86), 5ng/ml IL-4 (R&D #404-ML-010) and 0.5 ng/ml TGF-β1 (R&D #240-B-002), and grown for 72 h [22 (link)]. Cells were stained with FITC-conjugated anti-mouse IgA antibody (BD #559354) and analyzed by flow cytometry. CSR efficiency was determined by the percentage of IgA+ cells.

CH12F3.2a and its derivative cells were cultured in 10%FCS RPMI medium supplemented with 50 μM β-mercaptoethanol. As for CSR assay, healthy cells in log phase were seeded at 5×10⁴ cells/ml in medium with 1μg/ml anti-CD40 (eBioscience #16-0404-86), 5ng/ml IL-4 (R&D #404-ML-010) and 0.5ng/ml TGF- β1 (R&D #240-B-002), and grown for 72h. Cells were stained with FITC-conjugated anti-mouse IgA antibody (BD #559354) and analyzed by flow cytometry. CSR efficiency was determined by the percentage of IgA+ cells.