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Alexa Fluor® 647 Mouse Anti-Human Granzyme K
Alexa Fluor® 647 Mouse Anti-Human Granzyme K
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.
Product Details
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BD Pharmingen™
GZMK; GRAK; TRYP2; fragmentin-3; granzyme 3; tryptase II
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human Granzyme K Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2869812
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566655 Rev. 1
Antibody Details
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G3H69

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.

        

566655 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
566655 Rev.1
Citations & References
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Development References (5)

  1. Bade B, Boettcher HE, Lohrmann J, et al. Differential expression of the granzymes A, K and M and perforin in human peripheral blood lymphocytes.. Int Immunol. 2005; 17(11):1419-28. (Biology). View Reference
  2. Hameed A, Lowrey DM, Lichtenheld M, Podack ER. Characterization of three serine esterases isolated from human IL-2 activated killer cells. J Immunol. 1988; 141:3142-3147. (Biology). View Reference
  3. Inagaki H, Hirata Y, Shimizu T, Kobayashi M, Li Q, Kawada T. Expression of granzyme 3 protein in human peripheral blood lymphocytes analyzed by flow cytometry (PP-108-30). Int Immunol (Abstract). 2010; 22:172. (Immunogen: Flow cytometry).
  4. Lieberman J. The ABCs of granule-mediated cytotoxicity: new weapons in the arsenal.. Nat Rev Immunol. 2003; 3(5):361-70. (Biology). View Reference
  5. Wilharm E, Parry MA, Friebel R, et al. Generation of catalytically active granzyme K from Escherichia coli inclusion bodies and identification of efficient granzyme K inhibitors in human plasma. J Biol Chem. 1999; 274:27331-27337. (Biology). View Reference
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566655 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.