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RB780 Mouse Anti-Human Granzyme K
RB780 Mouse Anti-Human Granzyme K
Two-color flow cytometric analysis of Granzyme K expression in Human peripheral blood lymphocytes.  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 BD Horizon™ BV421 Mouse Anti-Human CD8 antibody (Cat. No. 562428) and with either BD Horizon™ RB780 Mouse IgG2a, κ Isotype Control (Cat. No. 568740; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human Granzyme K antibody (Cat. No. 569225/569226; Right Plot). The pseudocolor density plot showing the correlated expression of Granzyme K (or Ig Isotype control staining) versus CD8 was 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 lymphocytes.  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 BD Horizon™ BV421 Mouse Anti-Human CD8 antibody (Cat. No. 562428) and with either BD Horizon™ RB780 Mouse IgG2a, κ Isotype Control (Cat. No. 568740; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human Granzyme K antibody (Cat. No. 569225/569226; Right Plot). The pseudocolor density plot showing the correlated expression of Granzyme K (or Ig Isotype control staining) versus CD8 was 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.
商品详情
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BD Horizon™
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/test
3003
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


准备和存储

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 the dye under optimum conditions and unreacted dye was removed.

推荐的实验流程

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

商品通知

  1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  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. 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).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  11. For U.S. patents that may apply, see bd.com/patents.
569226 Rev. 2
抗体详情
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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.

569226 Rev. 2
格式详情
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RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
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RB780
Blue 488 nm
498 nm
781 nm
569226 Rev.2
报价单和参考
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研发参考 (6)

  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). 查看参考
  2. Baracho GV, Kara N, Rigaud S, Lo E, Widmann SJ, Tyznik AJ. Functional phenotyping of circulating human cytotoxic T cells and NK cells using a 16-color flow cytometry panel.. STAR Protoc. 2022; 3(1):101069. (Clone-specific: Cytotoxicity, Functional assay, Intracellular Staining/Flow Cytometry). 查看参考
  3. 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). 查看参考
  4. 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: Intracellular Staining/Flow Cytometry).
  5. Lieberman J. The ABCs of granule-mediated cytotoxicity: new weapons in the arsenal.. Nat Rev Immunol. 2003; 3(5):361-70. (Biology). 查看参考
  6. 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). 查看参考
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569226 Rev. 2

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