Recombinant Human g-Glutamylcyclotransferase/CRF21, CF

R&D Systems | Catalog # 5086-CT

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human g-Glutamylcyclotransferase/CRF21 (5086-CT)
  • 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

E. coli

Accession Number

O75223

Applications

Enzyme Activity
View all gamma-Glutamylcyclotransferase/CRF21 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human gamma-Glutamylcyclotransferase/CRF21 protein
Met1-Leu188

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

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

N-terminal Sequence Analysis

Ala2

Predicted Molecular Mass

21 kDa

SDS-PAGE

20-22 kDa, reducing conditions

Activity

Measured by its ability to release L-alanine from gamma -glutamyl-L-alanine, with the detection of alanine by alanine dehydrogenase.
The specific activity is >40,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5086-CT
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: gamma-Glutamylcyclotransferase/CRF21

Gamma-glutamylcyclotransferase (GGCT) is an essential enzyme in the gamma -glutamyl cycle that catalyzes the formation of 5-oxoproline from gamma-glutamyl dipeptides. The enzyme plays an important role in glutathione homeostasis. GGCT is also known as cytochrome c-releasing factor 21(CRF-21). It induces the release of cytochrome c from mitochondria, leading to apoptosis (1). It is a 188 amino acid protein that consists of six beta -strands, five alpha -helices and four short 3­­10 helices (2). Up-regulation of the expression of GGCT has been found in various tumor tissues including lung, esophagus, stomach, bile duct, uterine cervix, colon and breast. (3, 4). Therefore, GGCT is a potential biomarker for numerous types of cancers.

References

  1. Masuda Y. et al. (2006) Biochem. Biophys. Res. Commun. 346:454.
  2. Oakley A.J. et al. (2008) J. Biol. Chem. 283:22031.
  3. Gromov P. et al. (2010) J. Proteome Res. 9:3941.
  4. Amano T. et al. (2012) J. Histochem. Cytochem. 60:76.

Long Name

Cytochrome c Releasing Factor 21

Alternate Names

C7orf24, CRF21, gammaGlutamylcyclotransferase, GCTG, GGC, GGCT, LOC79017

Entrez Gene IDs

79017 (Human); 110175 (Mouse); 685702 (Rat)

Gene Symbol

GGCT

UniProt

O75223 (Human)

Additional gamma-Glutamylcyclotransferase/CRF21 Products

Product Documents for Recombinant Human g-Glutamylcyclotransferase/CRF21, 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 g-Glutamylcyclotransferase/CRF21, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Related Research Areas

Cancer Biomarkers

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Protocols

View specific protocols for Recombinant Human g-Glutamylcyclotransferase/CRF21, CF (5086-CT):

Materials
  • Assay Buffer: 50 mM Tris, pH 9.0
  • Recombinant Human gamma ‑Glutamylcyclotransferase/CRF21 (rhCRF21) (Catalog # 5086-CT)
  • gamma -L-Glutamyl-L-alanine (Sigma, Catalog # 483834), 100 mM stock in deionized water
  • beta -Nicotinamide adenine dinucleotide ( beta -NAD) (Sigma, Catalog # N6522), 100 mM stock in deionized water
  • L-Alanine Dehydrogenase (Sigma, Catalog # A7653)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhCRF21 to 0.05 ng/μL in Assay Buffer.
  2. Prepare a Substrate Mixture containing 10 mM gamma -L-Glutamyl-L-alanine, 2 mM beta -NAD, and 0.008 units/μL L‑Alanine Dehydrogenase in Assay Buffer.
  3. Load into a plate 50 μL of 0.05 ng/μL rhCRF21, and start the reaction by adding 50 μL of Substrate Mixture. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of Substrate Mixture.
  4. Read plate at a wavelength of 340 nm (bottom read) in kinetic mode for 5 minutes.
  5. 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 6220 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD.
  • hCRF-21: 0.0025 μg
  • gamma -L-Glutamyl-L-alanine: 5 mM
  • beta -NAD: 1 mM
  • L-Alanine Dehydrogenase: 0.4 units

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