Granzyme B Antibody [B18.1]

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 μ l FACS buffer (PBS, 5% Fetal calf serum, 0.02% azide) containing 1 μ g 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 μ g/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 μ g/mL. Proteins were visualized using a peroxidase-conjugated antibody to mouse IgG (SBA) and a chemiluminescence detection system.

Granzyme B and perforin can be used as predictive markers of acute rejection in heart transplantation: S. Legros-Maida, et al.; Eur. J. Immunol. 24, 229 (1994) .

Perforin and granzyme B expression is associated with severe acute rejection. Evidence for in situ localization in alveolar lymphocytes of lung-transplanted patients: M.V. Clement, et al.; Transplantation 57, 322 (1994) .

Granzyme B and perforin lytic proteins are expressed in CD34+ peripheral blood progenitor cells mobilized by chemotherapy and granulocyte colony-stimulating factor: C. Berthou, et al.; Blood 86, 3500 (1995) .

Expansion of a peripheral blood perforin+ CD8+ T-cell subset in long- term surviving lung transplanted patients: C. Berthou, et al.; Transplant. Proc. 28, 1964 (1996) .