beta 2 Microglobulin Antibody (YA935)(PBS only)

MedchemExpress Validation
Western blot analysis was performed on extracts from U937 (lane 1, 15 μg), Hela (lane 2, 15 μg), Raji (lane 3, 15 μg), Jurkat (lane 4, 15 μg), Mouse kidney (lane 5, 15 μg), and Rat kidney (lane 6, 15 μg) using beta 2 Microglobulin Rabbit mAb.Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight.The primary antibody (1:1000 dilution) and the loading control antibody (beta-Actin, HY-P83730, 1:20000 dilution) were incubated in 5% non-fat milk in TBST for 1 hour at 37°C.Goat Anti-Rabbit IgG-HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

MedChemExpress Validation
Indirect ELISA analysis of B2M was performed by coating wells of a 96-well plate with 100 µl per well of B2M antigen (HY-P7625) at a concentration of 2 µg/mL overnight at 4 °C. Wells of the plate were washed and blocked with 1% BSA in 1× PBST buffer. The plate was then incubated with 100 µl per well of B2M antibody, starting at an initial concentration of 10,000 ng/mL and serially diluted down to 0.05 ng/mL, for 2 hours at room temperature. The plate was washed and incubated with 100 µl per well of an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:10,000 for one hour at room temperature. Detection was performed using TMB substrate for 15 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid, and absorbances were read on a spectrophotometer at 450 nm.