Recombinant Mouse Granzyme B (carrier-free)

Pricing & Availability
Regulatory Status
RUO
Other Names
Granzyme 2, cytotoxic T-lymphocyte-associated serine esterase 1, GZMB, CCP1, Asp-ase
Granzyme_B_Mouse_Recom_BA_082814.jpg
The activity of recombinant mouse 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 >3000 pmol/min/µg.
  • Granzyme_B_Mouse_Recom_BA_082814.jpg
    The activity of recombinant mouse 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 >3000 pmol/min/µg.
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554802 10 µg $223.00
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554804 25 µg $440.00
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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 deficient of granzyme B 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 CD56+ CD8- 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 cells induces 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.

Technical data sheet

Product Details

Source
Mouse Granzyme B, amino acids (Lys17-Ser247) (Accession# NP_038570.1), was expressed with a C-terminal 10 His tag and a linker sequence in 293E cell line.
Molecular Mass
This 246 amino acid recombinant protein has a predicted molecular mass of approximately 27.5 kD. The protein migrates at approximately 37 kD in DTT-reducing conditions and approximately 37 kD in non-reducing conditions by SDS-PAGE. The predicted N-terminal amino acid is Lys.
Purity
>95%, as determined by Coomassie stained SDS-PAGE.
Formulation
0.22 µm filtered protein solution is in 20 mM HEPES, 150 mM NaCl, and pH 7.4.
Endotoxin Level
Less than 1.0 EU per µg of protein as determine by the LAL method.
Concentration
All sizes are bottled at 100 µg/mL.
Storage & Handling
Unopened vial can be stored at -70°C for six months. For maximum results, quick spin vial prior to opening. The recombinant protein could be aliquoted and stored at -20°C in a sterile buffer (20 mM HEPES, 150 mM NaCl, and pH 7.4). Avoid repeated freeze/thaw cycles.
Activity
Recombinant mouse Granzyme B supplied in its inactive form. Once the mouse Granzyme B is activated by active mouse Capthepsin C/DPPI, it is able to cleave the peptide substrate t-Butyloxycaronyl-Ala-Ala-ThioBenzyl ester (Boc-AAD-SBzl) in the presence of 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB)1,2, with an activity > 3000 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 verified 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.

Application References

(PubMed link indicates BioLegend citation)
  1. Edwards KM, et al. 1999. J. Biol. Chem 274:30468. (Granzyme B activation protocol)
  2. Smyth MJ, et al. 1995. J. Immunol. 154:6299. (Granzyme B activation protocol)

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 cells with ConA/ IL-2 and CD4+ T cells with anti CD3/CD28 or CD3/CD46.
Interaction
Targets of CTL and NK
Bioactivity
Mouse Granzyme B activated by 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 Weissmann C, 1987. J. Immunol. 139:250.
5. Trapani JA, et al. 1988. Proc. Natl. Acad. Sci. USA 85:6924.
6. Hiebert PR and Granville DJ, 2012. Trends. Mol. Med. 18:732.
7. Saito Y, et al. 2011. J. Cardiol. 57:141.

Gene ID
14939 View all products for this Gene ID
UniProt
View information about Granzyme B on UniProt.org
Go To Top Version: 3    Revision Date: 02/10/2015

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