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Two-color flow cytometric analysis of Granzyme K expression in human peripheral blood leucocytes. Human peripheral blood cells were treated (15 min; room temperature) with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to lyse erythrocytes and fix leucocytes. The leucocytes were washed and then stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with Alexa Fluor® 488 Mouse anti-Human CD8 antibody (Cat. No. 557696) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human Granzyme K antibody (Cat. No. 566655; Right Plot). Pseudocolor dot plots showing the correlated expression of Granzyme K (or Ig Isotype control staining) versus CD8 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
Two-color flow cytometric analysis of Granzyme K expression in human peripheral blood leucocytes. Human peripheral blood cells were treated (15 min; room temperature) with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to lyse erythrocytes and fix leucocytes. The leucocytes were washed and then stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with Alexa Fluor® 488 Mouse anti-Human CD8 antibody (Cat. No. 557696) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human Granzyme K antibody (Cat. No. 566655; Right Plot). Pseudocolor dot plots showing the correlated expression of Granzyme K (or Ig Isotype control staining) versus CD8 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
Two-color flow cytometric analysis of Granzyme K expression in human peripheral blood leucocytes. Human peripheral blood cells were treated (15 min; room temperature) with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to lyse erythrocytes and fix leucocytes. The leucocytes were washed and then stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with Alexa Fluor® 488 Mouse anti-Human CD8 antibody (Cat. No. 557696) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human Granzyme K antibody (Cat. No. 566655; Right Plot). Pseudocolor dot plots showing the correlated expression of Granzyme K (or Ig Isotype control staining) versus CD8 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
Two-color flow cytometric analysis of Granzyme K expression in human peripheral blood leucocytes. Human peripheral blood cells were treated (15 min; room temperature) with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to lyse erythrocytes and fix leucocytes. The leucocytes were washed and then stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with Alexa Fluor® 488 Mouse anti-Human CD8 antibody (Cat. No. 557696) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human Granzyme K antibody (Cat. No. 566655; Right Plot). Pseudocolor dot plots showing the correlated expression of Granzyme K (or Ig Isotype control staining) versus CD8 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
The G3H69 monoclonal antibody specifically recognizes Granzyme K which is also known as GRAK, Granzyme-3, Fragmentin-3, Tryptase II (TRYP2), or NK-tryptase-2 (NK-Tryp-2). This granule-associated enzyme is a serine protease that cleaves peptides after the basic residues lysine and arginine. Granzyme K is encoded by GZMK (granzyme K) that belongs to the Granzyme subfamily (Granzymes A, B, H, K, M) within the peptidase S1 family. Granzyme K is expressed in the granules of CD56-bright natural killer (NK) cells, NKT cells, γδ T cells, and cytotoxic CD4+ T cells and CD8+ T cells that can kill pathogen-infected or tumor cells. Granzyme K can activate the SET complex enabling DNase NM23-H1 to make single-stranded nicks in chromosomal DNA which ultimately contribute to target cell death. Granzyme K can also act on Bid and p53 to activate certain cell death pathways. Elevated levels of Granzyme K have been observed in the blood and bodily fluids from individuals under certain infectious or inflammatory conditions.
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