Granzyme B Antibody [B18.1]

Application Note:

Granzyme B antibody can be used for detection of human granzyme B by flow cytometry, immunocytochemistry, immunohistochemistry (frozen sections, paraffin sections), immunoprecipitation and western blot. Method: Permeabilized human T cells (5 x 105) were incubated on ice for 30 min in 50 μl FACS buffer (PBS, 5% Fetal calf serum, 0.02% azide) containing 1 μg of FITC- labeled B18.1 antibody. After washing in FACS buffer, cells were analyzed by flow cytometry. Method: Human T cells were plated onto polylysine treated glass slides, fixed and permeabilized in methanol at -20oC for 5 min, then in acetone at -20oC for 30 sec. After 3 washes in PBS, 0.1% BSA, slides were incubated for 1 hr at room temperature (RT) with 20 μg/mL of B18.4 antibody in PBS, 0.1% BSA. After rinsing in PBS, a FITC-conjugated anti-mouse IgG was added for 30 min., slides washed again in PBS and visualized using a fluorescence microscope. Method: : Recombinant granzyme B was resolved by SDS-PAGE under reducing conditions, transferred to nitrocellulose and probed with the monoclonal anti-granzyme B antibody B18.1 at 1 μg/mL. Proteins were visualized using a peroxidase-conjugated antibody to mouse IgG (SBA) and a chemiluminescence detection system.