Revision 1

#99695Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

25-35

Source/Isotype:

Rabbit IgG

UniProt ID:

#P20718

Entrez-Gene Id:

2999

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Granzyme H (E3H7W) Rabbit mAb recognizes endogenous levels of total Granzyme H protein. This antibody does not cross-react with human Granzyme A, B, or K proteins. This antibody does not cross-react with mouse Granzyme B proteins.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln195 of human Granzyme H protein.

Background

Granzymes are a family of serine proteases expressed by cytotoxic T lymphocytes and natural killer (NK) cells and are key components of immune responses to pathogens and transformed cells (1). Granzymes are synthesized as zymogens and are processed into mature enzymes by cleavage of a leader sequence. They are released by exocytosis in lysosome-like granules containing perforin, a membrane pore-forming protein. Granzyme B has the strongest apoptotic activity of all the granzymes as a result of its caspase-like ability to cleave substrates at aspartic acid residues thereby activating procaspases directly and cleaving downstream caspase substrates (2,3).
Granzyme H has chymotrypsin-like thioester activity with a preference for hydrophobic, aromatic amino acid residues, such as phenylalanine, tyrosine, or methionine, at the P1 site (4,5). Granzyme H is predominantly expressed at high levels in NK cells, but not in T lymphocytes, and has also been described in mast cells (6-8). After perforin-mediated entry into a target cell, Granzyme H induces many hallmarks of programmed cell death, such as mitochondrial depolarization, generation of reactive oxygen species, DNA degradation, and chromatin condensation (9,10).

  1. Trapani, J.A. (2001) Genome Biol. 2, REVIEWS 3014.
  2. Lord, S.J. et al. (2003) Immunol. Rev. 193, 31-8.
  3. Trapani, J.A. and Sutton, V.R. (2003) Curr. Opin. Immunol. 15, 533-43.
  4. Edwards, K.M. et al. (1999) J Biol Chem 274, 30468-73.
  5. Mahrus, S. and Craik, C.S. (2005) Chem Biol 12, 567-77.
  6. Sedelies, K.A. et al. (2004) J Biol Chem 279, 26581-7.
  7. Bade, B. et al. (2005) Int Immunol 17, 1419-28.
  8. Rönnberg, E. et al. (2014) Int Arch Allergy Immunol 165, 68-74.
  9. Fellows, E. et al. (2007) Blood 110, 544-52.
  10. Hou, Q. et al. (2008) Mol Immunol 45, 1044-55.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#99695

Granzyme H (E3H7W) Rabbit mAb

Western Blotting Image 1: Granzyme H (E3H7W) Rabbit mAb Expand Image
Western blot analysis of extracts from CD56+ NK cells, NK-92 cells, and SU-DHL-1 cells using Granzyme H (E3H7W) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: Granzyme H (E3H7W) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected (+) with constructs expressing Myc/DDK-tagged full-length human Granzyme A protein (hGranzyme A-Myc/DDK), Myc/DDK-tagged full-length human Granzyme B protein (hGranzyme B-Myc/DDK), Myc/DDK-tagged full-length mouse Granzyme B protein (mGranzyme B-Myc/DDK), Myc/DDK-tagged full-length human Granzyme H protein (hGranzyme H-Myc/DDK), and Myc/DDK-tagged full-length human Granzyme K protein (hGranzyme K-Myc/DDK), using Granzyme H (E3H7W) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Flow Cytometry Image 1: Granzyme H (E3H7W) Rabbit mAb Expand Image
Flow cytometric analysis of human peripheral blood mononuclear cells pre-gated on the lymphocyte population using Granzyme H (E3H7W) Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left), co-stained with CD3 (UCHT1) Mouse mAb (violetFluor 450 Conjugate) #61347. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.