Recombinant Human Transglutaminase 7/TGM7 Protein, CF

R&D Systems | Catalog # 5426-TG

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Transglutaminase 7/TGM7 Protein (5426-TG)
  • 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

Q96PF1

Applications

Enzyme Activity
View all Transglutaminase 7/TGM7 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Transglutaminase 7/TGM7 protein
Asp2-Pro710, with an N-terminal Met and 6-His tag
Accession # Q96PF1

Purity

>90%, 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

No result obtained

Predicted Molecular Mass

81 kDa

SDS-PAGE

75-78 kDa

Activity

Measured by its ability to form CBZ-Gln-Gly-Hydroxamate from CBZ-Gln-Gly and Hydroxylamine.
The specific activity is >350 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5426-TG
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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: Transglutaminase 7/TGM7

Transglutaminase 7 (TG7), encoded by the TGM7 gene, is also known as protein-glutamine-g-glutamyltransferase Z (Tgase Z) (1). It belongs to the family of Transglutaminases that catalyze the posttranslational modification of proteins via calcium dependent cross-linking reactions (2-4). TG7 is ubiquitously expressed in humans (1). Members of the TGM family have been implicated in a variety of human diseases including neurodegenerative diseases, celiac disease, lamellar ichthyosis, bleeding disorders, cataract formation, atherosclerosis, and others (5).

References

  1. Grenard, P. et al. (2001) J. Biol. Chem. 276:33066.
  2. Gentile, V. et al. (1991) J. Biol. Chem. 266:478.
  3. Chen, J.S.K. and Mehta K. (1999) Internat. J. Biochem. Cell Biol. 31:817.
  4. Griffin, M. et al. (2002) Biochem. J. 368:377.
  5. Kim, S-Y. et al. (2002) Neurochem. Int. 40:85.

Alternate Names

TGM7, TGMZ, TGZ

Entrez Gene IDs

116179 (Human); 640543 (Mouse); 691932 (Rat)

Gene Symbol

TGM7

UniProt

Q96PF1

Additional Transglutaminase 7/TGM7 Products

Product Documents for Recombinant Human Transglutaminase 7/TGM7 Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Transglutaminase 7/TGM7 Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human Transglutaminase 7/TGM7 Protein, CF (5426-TG):

Materials
  • Recombinant Human Transglutaminase 7/TGM7 (rhTGM7) (Catalog # 5426-TG)
  • Substrate: Z-Gln-Gly (Sigma, Catalog # C6154), 500 mM (dissolve in deionized water, then adjust to pH 9.0 with NaOH)
  • 0.1 M MES, pH 6.0
  • Dithiothreitol (DTT), 1 M stock in DMSO
  • 1.0 M CaCl2
  • 1.0 M Hydroxylamine Hydrochloride (Sigma, Catalog # 159417) (Dissolve in deionized water, then adjust to pH 6.0 with NaOH)
  • Stop Solution: 0.37 M FeCl3 (Sigma, Catalog # 236489), 0.67 M HCl, 0.2 M Trichloroacetic Acid
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Prepare substrate mixture right before running assay. Mix the following components per reaction:
    1. 15 µL 500 mM Z-Gln-Gly
    2. 75 µL 400 mM MES, pH 6.0
    3. 7.5 µL 200 mM DTT
    4. 7.5 µL 200 mM CaCl2
    5. 15 µL 1 M Hydroxylamine Hydrochloride

      6. Note: Multiply the volume for each component by the number of reaction vials + 1 to make enough substrate mixture for the assay. (For example: If there are 4 reaction vials, including blanks, multiply all volumes by 5)
  2. Dilute rhTGM7 to 0.1 mg/mL in deionized water.
  3. Mix 30 µL of the diluted rhTGM7 with 120 µL substrate mixture (step #1). Include a Substrate Blank containing 30 µL deionized water and 120 µL substrate mixture.
  4. Incubate at 37 °C for 2 hours.
  5. After incubation, stop the reaction with 600 µL of the Stop Solution. Mix well.
  6. Centrifuge at top speed for 2 minutes in a microcentrifuge.
  7. Transfer 200 µL (in duplicate) of the supernatant into a plate.
  8. Read at 525 nm (absorbance) in endpoint mode.
  9. Calculate specific activity:

     Specific Activity (pmoles/min/µg) =

 Adjusted Abs* (OD) x Conversion Factor** (pmole/OD)
Incubation time (min) x amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard L-glutamic acid g-monohydroxamate (Sigma, Catalog # G2253).

Per Well:
  • rhTGM7: 0.8 µg
  • Substrate: 10 mM

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