Recombinant Human Granzyme A (carrier-free)

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Other Names
Hanukah factor serine protease (HFSP), Cytolytic T cell-and natural killer cell-specific trypsin-like serine protease, Cytotoxic T-lymphocyte-associated serine esterase 3 (CTLA3)
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The activity of recombinant human granzyme A is determined by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Gly-Arg-ThioBenzyl ester (Z-GR-SBzl), in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB) with an activity of >5,000 pmol/min/µg.
  • Granzyme_A_Human_Recom_BA_043014.jpg
    The activity of recombinant human granzyme A is determined by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Gly-Arg-ThioBenzyl ester (Z-GR-SBzl), in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB) with an activity of >5,000 pmol/min/µg.
Cat # Size Price Quantity Avail. Save
550802 10 µg $185
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550804 25 µg $370
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Description

Granzyme A is a serine protease belonging to the granzyme family and is expressed exclusively by cytotoxic T cells (CTL) and NK cells. Most circulating CD56+CD8- NK cells, and approximately half of circulating CD8+ T cells, coexpress both granzymes A and B. In contrast, few circulating CD4+ T cells express granzyme A or B. Activation of CD8+ and CD4+ T lymphocytes induces substantial expression of granzyme B, but not granzyme A. Following receptor-mediated conjugate formation between a granzyme-containing cell and an infected or transformed target cell, granzymes enter the target cell via endocytosis and induce apoptosis. Granzyme A was found to induce caspase independent cell death when it enters into the target cell by perforin. Once in a cell, granzyme A activates DNA nicking by DNAse NM23-H1, a tumor suppressor gene product whose expression is reduced in transformed, metastatic cells. Dysregulation of this pathway results in several human diseases, such as hemophagocytic lymphohistiocytosis. Besides the protease activity, granzyme A induces human lung fibroblasts to produce IL-6 and IL-8. Cytokine induction is abrogated by treating the serine protease with the suicide serine protease inhibitor 3,4-dichloroisocoumarin. Other fibroblast lines, as well as epithelial cells, produce cytokines in response to granzyme A. These findings suggest that granzyme A can function as an activation molecule with potentially important immunoregulatory functions. However, CTLs from mice lacking granzyme A induce morphologically normal apoptosis in vitro, but those from mice that are deficient of granzyme B induce the nuclear features of apoptosis (particularly DNA fragmentation) more slowly than do wild-type CTLs.

Product Details
Technical data sheet

Product Details

Reactivity
Human
Source
Human Granzyme A, amino acids (Cys26-Val262) (Accession# NP_006135.1), is expressed with a C-terminal 10 His tag and a linker sequence in the 293E cell line.
Molecular Mass
This 252 amino acid recombinant protein has a predicted molecular mass of approximately 28 kD. The protein migrates at about 35 kD in DTT-reducing conditions and about 55 kD in non-reducing conditions by SDS-PAGE.The predicted N-terminal amino acid is Cys.
Purity
>90%, as determined by Coomassie stained SDS-PAGE.
Formulation
0.22 µm filtered protein solution is in 20 mM Tris, 150 mM NaCl, pH 7.5.
Endotoxin Level
Less than 1.0 EU per µg of protein as determine by the LAL method.
Concentration
10 and 25 µg sizes are bottled at 200 µg/mL.
Storage & Handling
Unopened vial can be stored at -70°C for six months. For maximum results, quick spin vial prior to opening. Avoid repeated freeze/thaw cycles.
Activity
Lysyl endopeptidase activated human granzyme A cleaves the peptide substrate N-carbobenzyloxy-Gly-Arg-ThioBenzyl ester (Z-GR-SBzl), in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB), with an activity >5,000 pmol/min/µg.
Application

Bioassay

Application Notes

This protein is in the latent form and needs to be activated for bioassay.


BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.

Antigen Details

Structure
Monomer
Distribution

CTL and NK cells.

Function
Granzyme A is able to induce target cell apoptosis by activating caspase independent pathways.
Interaction
Targets of CTL and NK cells.
Bioactivity
Lysyl endopeptidase activated human granzyme A is able to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Gly-Arg-ThioBenzyl ester (Z-GR-SBzl), in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB).
Biology Area
Cell Biology, Immunology, Innate Immunity, Neuroscience
Molecular Family
Proteases, Enzymes and Regulators
Antigen References

1. Edwards KM, et al. 1999. J. Biol. Chem. 274:30468.
2. Lieberman J, and Fan Z. 2003. Curr. Opin. Immunol. 15:553.
3. Fan Z, et al. 2003. Cell. 112:659.
4. Sower LE, et al. 1996. Cell Immunol. 171:159.
5. Grossman WJ. et al. 2004. Blood. 104:2840.
6. Ebnet K, et al. 1995. EMBO J. 14:4230.
7. Heusel JW, et al. 1994. Cell. 76:977.

Gene ID
3001 View all products for this Gene ID
UniProt
View information about Granzyme A on UniProt.org

Related FAQs

Does specific activity of a recombinant protein vary between lots?

Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.

Have your recombinants been tested for stability?

Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.

How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
How does the activity of your recombinant proteins compare to competitors?

We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!

What is the specific activity or ED50 of my recombinant protein?

The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.

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For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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