Recombinant Human Granzyme A Protein, CF

R&D Systems | Catalog # 2905-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Granzyme A Protein (2905-SE)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

P12544

Structure / Form

Pro form

Applications

Enzyme Activity
View all Granzyme A Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Granzyme A protein
Cys26-Val262, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Cys26

Predicted Molecular Mass

28 kDa

SDS-PAGE

Multiple bands between 29-33 kDa, reducing conditions

Activity

Measured 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). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468.
The specific activity is >5,000 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2905-SE
Formulation Lyophilized from a 0.2 μm filtered solution in MES, NaCl and CaCl2 with Trehalose.
Reconstitution Reconstitute at 100 μg/mL in sterile 25 mM HEPES, 150 mM NaCl and 10 mM CaCl2, pH 7.5.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Granzyme A

Granzyme A is a member of the granzyme family of the serine proteases found specifically in the cytotoxic granules of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. Granzyme A is the most abundant protease in CTL and NK cells. It induces caspase‑independent cell death when introduced into target cells by perforin (1). Human granzyme A is synthesized as a precursor (262 residues) with a signal peptide (residues 1‑26), a propeptide (residues 27-28) and a mature chain (residues 29-262 ) (2). The purified recombinant human Granzyme A consists of residues 26 to 262. After being activated by lysyl endopeptidase, it cleaves a thioester substrate as described in Activity Assay Protocol.

References

  1. Lieberman, J. and Z. Fan (2003) Curr. Opin. Immunol. 15:553.
  2. Gershenfeld, H.K. et al. (1988) Proc. Natl. Acad. Sci. USA 85:1184.

Alternate Names

CTL Tryptase, CTLA3, Fragmentin-1, GZMA, HF, HFSP

Entrez Gene IDs

3001 (Human); 14938 (Mouse); 266708 (Rat)

Gene Symbol

GZMA

UniProt

P12544 (Human)

Additional Granzyme A Products

Product Documents for Recombinant Human Granzyme A Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Granzyme A Protein, CF

For research use only

Citations for Recombinant Human Granzyme A Protein, CF

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Protocols

View specific protocols for Recombinant Human Granzyme A Protein, CF (2905-SE):

Materials
  • Activation Buffer: 0.1 M Tris, pH 9.0
  • Assay Buffer: 50 mM Tris, pH 8.0
  • Recombinant Human Granzyme A (rhGranzyme A) (Catalog # 2905-SE)
  • Lysyl-Endopeptidase
  • Substrate: Z-Gly-Arg-thiobenzyl ester, 10 mM stock in DMSO
  • 5,5'Dithio-bis(2-nitrobenzoic acid) (DTNB), 10 mM stock in DMSO
  • Clear 96-well Plate
  • Plate Reader with Absorbance Read Capability
  1. Activate rhGranzyme A at 50 µg/mL with 0.1 µg/mL Lysyl-Endopeptidase in Activation Buffer.
  2. Incubate at 37 °C for 1 hour.
  3. Dilute activated rhGranzyme A to 0.2 ng/µL in Assay Buffer.
  4. Dilute Substrate to 200 µM in Assay Buffer containing 200 µM of DTNB.
  5. In a plate, load 50 µL of 0.2 ng/µL rhGranzyme A, and start the reaction by adding 50 µL of 200 µM Substrate mixture. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of Substrate mixture.
  6. Read absorbance at a wavelength of 405 nm, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
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 0.320 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rhGranzyme A: 0.01 µg
  • DTNB: 100 µM
  • Substrate: 100 µM

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