Recombinant Mouse Granzyme G Protein, CF

R&D Systems | Catalog # 1346-SE

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Mouse Granzyme G Protein (1346-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

Sf 21 (baculovirus)

Accession Number

P13366

Structure / Form

Mature form

Applications

Enzyme Activity
View all Granzyme G Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Granzyme G protein
Ile21-Leu248, with a C-terminal 10-His tag
Accession # P13366

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

N-terminal Sequence Analysis

Ile21

Predicted Molecular Mass

27 kDa

SDS-PAGE

35 kDa, reducing conditions

Activity

Measured by its ability to cleave a peptide substrate, Succinyl-Phe-Leu-Phe-ThioBenzyl ester (Suc-FLF-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 >150 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

1346-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. 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 opening.

Background: Granzyme G

Granzyme G 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 (1). Together with Granzymes D, E and F, it is regulated through pregnancy and by IL-2 and IL-15 in granulated metrial gland cells (2). Human or rat counterpart of mouse Granzyme G has not been found. Like other Granzymes, mouse Granzyme G is not secreted as a zymogen but stored as a fully processed and activated enzyme in the cytoplasmic granules of CTL (3). It is synthesized as a 248 amino acid precursor with a 18 amino acid signal peptide and a 2 amino acid propeptide (3, 4). The mature protein (residues 21‑248) is expressed and purified.

References

  1. Smyth, M.J. et al. (1996) J. Leukoc. Biol. 60:555.
  2. Allen, M.P. and M. Nilsen-Hamilton (1998) J. Immunol. 161:2772.
  3. Jenne, D.E. et al. (1989) Biochemistry 28:7953.
  4. Kwon, B.S. et al. (1988) J. Exp. Med. 168:1839.

Alternate Names

GrzG, GZMG

Entrez Gene IDs

14944 (Mouse); 266704 (Rat)

Gene Symbol

GZMG

UniProt

P13366

Additional Granzyme G Products

Product Documents for Recombinant Mouse Granzyme G Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Mouse Granzyme G Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Mouse Granzyme G Protein, CF (1346-SE):

Materials
  • Assay Buffer: 0.1 M Tris, pH 8.0
  • Recombinant Mouse Granzyme G (rmGranzyme G) (Catalog # 1346-SE)
  • Substrate: SUC-Phe-Leu-Phe-SBZL (Bachem, Catalog # M-1740), 10 mM in DMSO
  • 5,5’-dithio-bis (2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM in DMSO
  • 96 well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rmGranzyme G to 2 ng/µL in Assay Buffer.
  2. Combine equal volumes of Substrate and DTNB for a final concentration of 5 mM of each.
  3. Dilute the Substrate/DTNB mixture to 200 µM in Assay Buffer.
  4. Load 50 µL of the diluted rmGranzyme G into a 96 well clear plate. Include a Substrate Blank containing 50 µL of Assay Buffer.
  5. Start the reaction by adding 50 µL of the diluted Substrate/DTNB mixture to wells.
  6. Read at a wavelength of 405 nm (bottom read) 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 0.32 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rmGranzyme G: 0.1 µg
  • Substrate: 100 µM
  • TNB: 100 µM

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