Intracellular Staining by Flow Cytometry
|Detection of Granzyme B in NK‑92 Human Cell Line by Flow Cytometry. NK‑92 human natural killer lymphoma cell line was stained with Mouse Anti-Human Granzyme B PE‑conjugated Monoclonal Antibody (Catalog # IC2906P, filled histogram) or isotype control antibody (Catalog # IC003P, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.|
Granzyme B is a member of the Granzyme family of the serine proteases found specifically in the cytotoxic granules of Cytotoxic T Lymphocytes (CTL) and Natural Killer (NK) cells (1, 2). Granzyme B plays an essential role in granule-mediated apoptosis and may have additional roles in rheumatoid arthritis and in bacterial and viral infections (3). It activates various Caspases and cleaves proteins such as Aggrecan (3). Human Granzyme B is synthesized as a 247 amino acid (aa) precursor with a signal peptide (aa 1-18), a pro peptide (aa 19-20), and a mature chain (aa 21-247) (4-6). The recombinant human (rh) Granzyme B consisting of aa 19-247 was expressed and purified. After being activated by active Cathepsin C, rhGranzyme B cleaves a thioester substrate described previously (3).
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