Recombinant Human Granzyme B (carrier-free)

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Other Names
Granzyme 2, cytotoxic T-lymphocyte-associated serine esterase 1, GZMB, CCP1, Asp-ase
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Granzyme_B_Human_Recom_BA_081914.jpg
The activity of recombinant human Granzyme B is determined by its ability to cleave a colorimetric peptide substrate, t-Butyloxycaronyl-Ala-Ala-Asp-ThioBenzyl ester (Boc-AAD-SBzl), in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB), with an activity >1500 pmol/min/µg.
  • Granzyme_B_Human_Recom_BA_081914.jpg
    The activity of recombinant human Granzyme B is determined by its ability to cleave a colorimetric peptide substrate, t-Butyloxycaronyl-Ala-Ala-Asp-ThioBenzyl ester (Boc-AAD-SBzl), in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB), with an activity >1500 pmol/min/µg.
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Cat # Size Price Quantity Avail. Save
554902 10 µg $190
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554904 25 µg $375
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Description

Granzyme B is a serine protease expressed by cytotoxic T cells (CTL) and NK cells. Its main function is to induce cell death to eliminate harmful targets such as allogeneic, virally infected, and tumorous cells. This is evident by the fact that CTLs from mice with inhibited granzyme B production exhibit a profound defect in inducing rapid DNA fragmentation and apoptosis in target cells. Following receptor-mediated conjugate formation between CTL or NK and their target cell, granzyme B enters the target via endocytosis, and subsequently activates multiple protein substrates to induce apoptosis. Most circulating CD56CD8- NK cells, and approximately half of circulating CD8+ T cells, coexpress both granzyme A and B. In contrast, few circulating CD4+ T cells express granzymes A or B. Activation of CD8+ and CD4+ T lymphocytes induce substantial expression of granzyme B, but not granzyme A. Besides CTL and NK, evidence has shown that the distribution of human granzyme B has a broader spectrum of cells, including CD34+ hematopoietic progenitor cells, keratinocytes, basophils, mast cells, plasmacytoid dendritic cells, and B cells. Although its role in cytotoxic lymphocyte-mediated apoptosis is well established, granzyme B can also degrade extracellular matrix proteins and alter inflammation if present in the extracellular milieu. These findings suggest that granzyme B can function as an activation molecule with potentially important immunoregulatory functions. In addition, it was shown that expression of granzyme B is elevated in acute coronary syndrome and acute myocardia infarction, indicating that granzyme B could be a factor involved in cardiovascular diseases.

Product Details
Technical data sheet

Product Details

Reactivity
Human
Source
Human Granzyme B, amino acids (Gly19-Tyr249) (Accession# NP_004122.2), was expressed with a C-terminal 10 His tag and a linker sequence in 293E cell line.
Molecular Mass
This 244 amino acid recombinant protein has a predicted molecular mass of approximately 27.5 kD. The protein migrates at about 37 kD in DTT-reducing conditions and about 37 kD in non-reducing conditions by SDS-PAGE.The predicted N-terminal amino acid is Gly.
Purity
>95%, 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 - 100 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
Human Granzyme B activated by mouse Cathepsin C is able to cleave the peptide substrate t-Butyloxycaronyl-Ala-Ala-Asp-ThioBenzyl ester (Boc-AAD-SBzl), in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB), with an activity >1500 pmol/min/µg. This recombinant protein is in its inactive form and needs to be activated prior to use in bioassays. Please contact technical support for a detailed activation protocol.
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

Cytotoxic T cells, NK cells, and neutrophils.

Function
Granzyme B is able to induce target cell apoptosis by activating caspase independent pathways. Granzyme B is induced in CD8+ T lymphocytes with ConA / IL-2 and CD4+ T lymphocytes with anti CD3/CD28 or CD3/CD46.
Interaction
Targets of CTL and NK cells.
Bioactivity
Human Granzyme B activated by mouse Cathepsin C is able to cleave the peptide substrate t-Butyloxycaronyl-Ala-Ala-Asp-ThioBenzyl ester (Boc-AAD-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. Grossman WJ, et al. 2004. Blood. 104:2840.
3. Heusel JW, et al. 1994. Cell. 76:977.
4. Schmid J and Weissman C. 1987. J. Immunol. 139:250.
5. Trapani JA, et al. 1988. Proc. Natl. Acad. Sci. USA 85:6924.
6. Heibert PR, and Granville DJ. 2012. Trends. Mol. Med. 18:732.
7. Saito Y, et al. 2011. J. Cardiol. 57:141.

Gene ID
3002 View all products for this Gene ID
UniProt
View information about Granzyme B 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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