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Figure: Detection of granzyme B in a human CD8+ T cell clone.
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.
Figure: Detection of recombinant granzyme B. Granzyme B migrates as a 27 kDa species.
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 1µg/ml. Proteins were visualized using a peroxidase-conjugated antibody to mouse IgG (SBA) and a chemiluminescence detection system.
Schematic structure of human granzyme B (recombinant, aa. 7-227).
| Regulatory Status |
RUO – Research Use Only |
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