Granzyme K Monoclonal Antibody (G3H69), PerCP-eFluor™ 710, eBioscience™

Product Specific Information

Description: This G3H69 monoclonal antibody reacts with human Granzyme K. Granzyme K is one of five granzyme serine proteases that have been identified in humans. These proteins are expressed in the granules of NK cells and cytotoxic T cells, and are critical for the induction of target cell apoptosis through the cleavage of intracellular substrates. Granzyme A and Granzyme K are both tryptases and appear to show overlapping, though not identical, substrate specificity. This functional similarity is believed to account for the minimal decrease in cytotoxicity of Granzyme A-deficient CTLs.

Applications Reported: This G3H69 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This G3H69 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood monocytes. This can be used at 5 µL (0.03 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Granzyme K (GrK) belongs to a family of trypsin-like serine proteases localised in the cytotoxic granules of activated T cells and NK cells. It encodes a 28 kDa serine protease whose gene is located on chromosome 5q11-12 close to the granzyme A-encoding gene. Like granzyme A, it has a trypsin-like specifity cleaving at the basic residues arginine and lysine. Granzyme K triggers rapid cell death independently of caspase activation with single-stranded DNA nicks and is primarily expressed in thymus, lung, spleen and peripheral blood leukocytes.