APRIL Antibody [Sacha-1]

Application Note:

APRIL antibody can be used to recognize human and mouse APRIL by Flow Cytometry (excellent for mouse APRIL, also stains human APRIL), and Immunocytochemistry. Method: 3 days after transfection of cells with the indicated constructs, cells were fixed with acetone. Slides were blocked with IgG, and incubated for 1 hour with 5 μg/mL Sacha-1 or control murine IgG (isotype control) in 1%BSA / 1x PBS for 1 hour. After washes wish PBS, samples were incubated with the secondary Ab for 1 hour, washed in PBS and revealed with StreptABComplex/HRP (Vector) and AEC. Method: 3 HEK 293T cells (5 x 105) were mock transfected (thin line) or transfected with an expression plasmid enabling surface expression of mAPRIL (thick line). Cells were incubated on ice for 30 min in 50 μl FACS buffer (PBS, 5% Fetal calf serum, 0.02% azide) containing 1 μg/mL of Sacha-1 antibody. After washing in FACS buffer, PE-conjugated antibody to rat IgG was added. Cells were incubated on ice for 30 min, washed and then analyzed by flow cytometry.