FITC Mouse Anti-Human Granzyme A

BD Pharmingen™ FITC Mouse Anti-Human Granzyme A

Name: FITC Mouse Anti-Human Granzyme A
Brand: FITC Mouse Anti-Human Granzyme A
SKU: 568661

Clone CB9 (RUO)

Multiparameter flow cytometric analysis of Granzyme A expression in Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) and with either FITC Mouse IgG1 Isotype Control (Cat. No. 340755; Left Plot) or FITC Mouse Anti-Human Granzyme A antibody (Cat. No. 568661/568662; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of Granzyme A (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

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Product Details

Brand: BD Pharmingen™

Alternative Name: CTL tryptase; CTLA3; GZMA; GrA; HFSP; Hanukkah factor

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human Cytotoxic T Cell Granule Proteins

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 5 µl

Entrez Gene ID: 3001

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: 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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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).
An isotype control should be used at the same concentration as the antibody of interest.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Mouse IgG1 FITC ASR

Size 100 Tests Cat No. 340755

Fixation Buffer RUO

Size 100 mL Cat No. 554655

Perm/Wash Buffer RUO

Size 100 mL Cat No. 554723

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

View Details

568661 Rev. 1

Antibody Details

CB9

The CB9 monoclonal antibody specifically recognizes Granzyme A (GrA, GzmA or Gzm A) which is also known as CTL tryptase,

Cytotoxic T-lymphocyte-associated serine esterase-3 (CTLA3), Cytotoxic T-lymphocyte proteinase 1, Fragmentin-1, or Hanukah factor serine protease (HFSP). Granzyme A belongs to a family of effector proteases that include five different granzymes that play key roles in innate and adaptive immune responses to pathogens and transformed cells: Granzymes A, B, H, K and M. Granzyme A is a ~28 kDa neutral serine protease that is encoded by GZMA. It is expressed in the cytotoxic granules of natural killer (NK) cells, NKT cells, cytotoxic αβ T cells and γδ T cells. Granzyme A participates in innate and adaptive immune responses. After gaining access into target cells, Granzyme A cleaves various peptide substrates after lysine or arginine residues and can induce caspase‑independent cell death reflected in single-stranded DNA damage as well as rapid loss of cell membrane integrity and mitochondrial transmembrane potential.

568661 Rev. 1

Format Details

FITC

Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: FITC

Excitation Source: Blue 488 nm

Excitation Max: 494 nm

Emission Max: 518 nm

568661 Rev.1

Citations & References

View product citations for antibody "568661" on CiteAb

Development References (8)

Andrade F, Roy S, Nicholson D, Thornberry N, Rosen A, Casciola-Rosen L. Granzyme B directly and efficiently cleaves several downstream caspase substrates: implications for CTL-induced apoptosis. Immunity. 1998; 8(4):451-460. (Biology). View Reference
Barry M, Heibein JA, Pinkoski MJ, et al. Granzyme B short-circuits the need for caspase 8 activity during granule-mediated cytotoxic T-lymphocyte killing by directly cleaving Bid. Mol Cell Biol. 2000; 20(11):3781-3794. (Biology). View Reference
Beresford PJ, Kam CM, Powers JC, Lieberman J. Recombinant human granzyme A binds to two putative HLA-associated proteins and cleaves one of them. Proc Natl Acad Sci U S A. 1997; 94(17):9285-9290. (Immunogen: Immunoprecipitation). View Reference
Beresford PJ, Xia Z, Greenberg AH, Lieberman J. Granzyme A loading induces rapid cytolysis and a novel form of DNA damage independently of caspase activation. Immunity. 1999; 10(5):585-594. (Clone-specific: Immunoprecipitation). View Reference
Pinkoski MJ, Waterhouse NJ, Heibein JA, et al. Granzyme B-mediated apoptosis proceeds predominantly through a Bcl-2-inhibitable mitochondrial pathway. J Biol Chem. 2001; 276(15):12060-12067. (Biology). View Reference
Sharif-Askari E, Alam A, Rheaume E, et al. Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation. EMBO J. 2001; 20(12):3101-3113. (Biology). View Reference
Shresta S, Graubert TA, Thomas DA, Raptis SZ, Ley TJ. Granzyme A initiates an alternative pathway for granule-mediated apoptosis. Immunity. 1999; 10(5):595-605. (Biology). View Reference
Trimble LA, Lieberman J. Circulating CD8 T lymphocytes in human immunodeficiency virus-infected individuals have impaired function and downmodulate CD3 zeta, the signaling chain of the T-cell receptor complex. Blood. 1998; 91(2):585-594. (Clone-specific: Flow cytometry). View Reference

View All (8)

568661 Rev. 1

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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.