Recombinant Human Transglutaminase 1/TGM1 Protein, CF

R&D Systems | Catalog # 7765-TG

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Transglutaminase 1/TGM1 Protein (7765-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

P22735

Applications

Enzyme Activity
View all Transglutaminase 1/TGM1 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Transglutaminase 1/TGM1 protein
Met2-Ala817, with an N-terminal Met and 6-His tag

Purity

>70%, 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

Inconclusive. Gly6, Ser7, Thr491, Ala571 identified. Poly His detected by Western blot.

Predicted Molecular Mass

89 kDa

SDS-PAGE

75-88 kDa, reducing conditions

Activity

Measured by its ability to form CBZ-Gln-Gly-Hydroxamate from CBZ-Gln-Gly and Hydroxylamine.
The specific activity is >100 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7765-TG
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, EDTA, DTT 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 1/TGM1

Transglutaminase 1, also known as TGM1, Transglutaminase K, keratinocyte transglutaminase, or epidermal TGase, is a member of the transglutaminase family. Transglutaminases catalyze the calcium-dependent formation of isopeptide bonds between lysine and glutamine residues (1). TGM1 plays an important role in formation of the cornified envelope of differentiating keratinocytes in the epidermis through its cross-linking of epidermal proteins (2). The majority of cases of lamellar ichthyosis result from mutations present within the TGM1 gene and the subsequent dysfunction of TGM1 (1, 3).

References

  1. Lorand, L. and R.M. Graham (2003) Nature Rev. Mol. Cell. Biol. 4:140.
  2. Eckert, R.L. et al. (2005) J. Invest. Dermatol. 124:481.
  3. Farasat, S. et al. (2009) J. Med. Genet. 46:103.

Alternate Names

ICR2, KTG, LI1, TGase-1, TGK, TGM1

Entrez Gene IDs

7051 (Human); 21816 (Mouse); 60335 (Rat)

Gene Symbol

TGM1

UniProt

P22735 (Human)

Additional Transglutaminase 1/TGM1 Products

Product Documents for Recombinant Human Transglutaminase 1/TGM1 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 Transglutaminase 1/TGM1 Protein, CF

For research use only

Related Research Areas

Extracellular Matrix Molecules

Citations for Recombinant Human Transglutaminase 1/TGM1 Protein, CF

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Protocols

View specific protocols for Recombinant Human Transglutaminase 1/TGM1 Protein, CF (7765-TG):

Materials
  • Assay Buffer: deionized water
  • Recombinant Human Transglutaminase 1/TGM1 (rhTGM1) (Catalog # 7765-TG)
  • Substrate: Z-Gln-Gly (Sigma, Catalog # C6154), 500 mM, pH 9.0 in deionized water
  • Hydroxylamine Hydrochloride (Sigma, Catalog # 159417), 1 M, pH 6.0 in deionized water
  • 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 containing 62.5 mM Substrate, 250 mM MES pH 6.0, 12.5 mM DTT, 12.5 mM CaCl2, and 125 mM Hydroxylamine Hydrochloride.
  2. Dilute rhTGM1 to 0.2 mg/mL in Assay Buffer.
  3. Mix 30 µL dilute rhTGM1 with 120 µL Substrate Mixture. Include an Enzyme Control containing 30 µL Assay Buffer with 120 µL Substrate Mixture.
  4. Incubate at 37 °C for 2 hours.
  5. Stop each reaction with 600 µL of Stop Solution. Mix well.
  6. Centrifuge at top speed for 2 minutes in a microcentrifuge.
  7. Load 200 µL of the supernatant into a 96 well clear plate.
  8. Read plate at 525 nm (absorbance) in endpoint mode.
  9. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

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

     *Adjusted for Enzyme Control
     **Derived using calibration standard L-glutamic acid gamma -monohydroxamate (Sigma, Catalog # G2253).

Per Well:
  • rhTGM1: 1.6 µg
  • Substrate: 10 mM

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