Recombinant Human Granzyme H Protein, CF Summary
Glu19-Leu246, with a C-terminal 10-His tag
Accession # P20718
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Supplied as a 0.2 μm filtered solution in HEPES, NaCl and Brij-35.|
|Shipping||The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- Activation Buffer: 50 mM MES, 50 mM NaCl, pH 5.5
- Assay Buffer: 50 mM Tris, 0.5 M NaCl, pH 9.0
- Recombinant Human Granzyme H (rhGranzyme H) (Catalog # 1377-SE)
- Recombinant Mouse Active Cathepsin C/DPPI (rmCathepsin C) (Catalog # 2336-CY)
- Substrate: Succinyl-Phe-Leu-Phe ThioBenzyl ester (Suc-FLF-SBzl) (Bachem, Catalog # M-1740), 10 mM stock in DMSO
- 5,5’-dithio-bis (2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM stock in DMSO
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Activate rhGranzyme H at 100 µg/mL with 21 µg/mL of rmCathepsin C in Activation Buffer.
- Incubate reaction at 37 °C for 2 hours.
- Dilute activated rhGranzyme H to 0.4 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM containing 200 µM of DTNB in Assay Buffer.
- In a plate load 50 µL of 0.4 ng/µL rhGranzyme H, and start the reaction by adding 50 µL of 200 µM Substrate mixture to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate mixture.
- Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
- Calculate specific activity:
Specific Activity (pmol/min/µg) =
|Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/M|
|ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)|
*Adjusted for Substrate Blank
**Using the extinction coefficient 13260 M-1cm-1
***Using the path correction 0.32 cm
Note: the output of many spectrophotometers is in mOD Per Well:
- rhGranzyme H: 0.02 µg
- DTNB: 100 µM
- Substrate: 100 µM
Background: Granzyme H
Granzyme H is a member of the granzyme family of serine proteases found specifically in the cytotoxic granules of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells (1, 2). Granzyme H’s functions are largely unknown. The more abundant expression of Granzyme H than Granzyme B in NK cells suggests that Granzyme H may complement the pro-apoptotic function of Granzyme B in this cell type (3). Human Granzyme H shows the highest amino acid identity (71%) to mouse Granzyme C (4). Human Granzyme H is synthesized as a precursor (246 residues) with a signal peptide (residues 1-18), a propeptide (residues 19-20) and a mature chain (residues 21-246) (5-7). The pro-enzyme is expressed and purified. After being activated by active cathepsin C, rhGranzyme H cleaves a thioester substrate, which was described previously (8).
- Kam, C.-M. et al. (2000) Biochim. Biophys. Acta 1477:307.
- Smyth, M.J. et al. (1996) J. Leukoc. Biol. 60:555.
- Sedelies, K.A. et al. (2004) J. Biol. Chem. 279:26581.
- Sattar, R. et al. (2003) Biochem. Biophys. Res. Comm. 308:726.
- Meier, M. et al. (1990) Biochemistry 29:4042.
- Haddad, P. et al. (1991) Int. Immunol. 3:57.
- Klein, J.L. et al. (1990) Tissue Antigens 35:220.
- Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468.
Citations for Recombinant Human Granzyme H Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 2
Filter your results:
Granzyme B PET Imaging as a Predictive Biomarker of Immunotherapy Response
Authors: BM Larimer, E Wehrenberg, F Dubois, A Mehta, T Kalomeris, K Flaherty, G Boland, U Mahmood
Cancer Res., 2017;77(9):2318-2327.
Sample Types: Protein
Applications: Enzyme Assay
Natural killer cell-derived human granzyme H induces an alternative, caspase-independent cell-death program.
Authors: Fellows E, Gil-Parrado S, Jenne D, Kurschus F
Applications: Enzyme Assay
No product specific FAQs exist for this product, however you mayView all Proteins and Enzyme FAQs
Reviews for Recombinant Human Granzyme H Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Granzyme H Protein, CF and earn rewards!
Have you used Recombinant Human Granzyme H Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image