Recombinant Human Transglutaminase 2/TGM2 Protein, CF

Catalog # Availability Size / Price Qty
4376-TG-050
R&D Systems Recombinant Proteins and Enzymes
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Citations (6)
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Recombinant Human Transglutaminase 2/TGM2 Protein, CF Summary

Product Specifications

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.
Activity
Measured by its ability to cleave a synthetic peptide Benzyloxycarbonyl-Gln-Gly and NH2OH. The specific activity is >1000 pmol/min/ug, as measured under the described condition.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Transglutaminase 2/TGM2 protein
Ala2-Ala687 (Asn533Thr and Leu655Val), with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Analysis
No results obtained
Predicted Molecular Mass
78 kDa
SDS-PAGE
82 kDa, reducing conditions

Product Datasheets

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4376-TG

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

4376-TG

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT.
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.

Assay Procedure

Materials
  • Assay Diluent: deionized water
  • Recombinant Human Transglutaminase 2/TGM2 (rhTGM2) (Catalog # 4376-TG)
  • Substrate: Z-Gln-Gly (Sigma, Catalog # C6154), dissolve 500 mM in deionized water, then neutralize it to pH 9.0 with NaOH
  • MES, pH 6.0, 1 M stock
  • DTT (Sigma, Catalog # D-0632), 1 M stock in deionized water
  • CaCl2,1 M stock in deionized water
  • 1 M Hydroxylamine Hydrochloride (Sigma, Catalog # 159417), dissolve in deionized H2O, then neutralize it 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 fresh before running assay. Mix the following components (Dilute all components to the correct concentration in Assay Diluent.):
    1. 10 µL of 500 mM Substrate
    2. 50 µL of 400 mM MES, pH 6.0
    3. 5 µL of 200 mM DTT
    4. 5 µL of 200 mM CaCl2
    5. 10 µL of 1 M Hydroxylamine Hydrochloride
      (Note: Multiply the volume for each component by the number of reaction vials + 1 to make enough substrate mixture for the assays. For example: If there are 2 reaction vials (including blank), multiply all volumes by 3.)
  2. Dilute rhTGM2 to 0.1 mg/mL in Assay Diluent.
  3. Mix 20 µL of the diluted rhTGM2 with 80 µL Substrate Mixture. For the Substrate Blank mix 20 µL of Assay Diluent with 80 µL Substrate Mixture.
  4. Incubate at 37 °C for 2 hours.
  5. After incubation, stop the reaction with 400 µL of the Stop Solution. Mix well.
  6. Centrifuge at top speed for 2 minutes in a microcentrifuge.
  7. Load 200 µL of the supernatant into a plate.
  8. Read 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 Substrate Blank
     **Derived using calibration standard L-glutamic acid gamma -monohydroxamate (Sigma, Catalog # G2253).

Per Well:
  • rhTGM2: 0.8 µg
  • Substrate: 10 mM
Reconstitution Calculator

Reconstitution Calculator

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: Transglutaminase 2/TGM2

Transglutaminase 2 (TG2), encoded by the TGM2 gene, is also known as tissue transglutaminase (tTG), transgluytaminase C (TGC), and protein‑glutamine‑ gamma -glutamyltransferase. It belongs to the family of transglutaminases that catalyze the posttranslational modification of proteins via calcium dependent cross-linking reactions (1‑3). In addition to its function in protein cross-linking, TGM2 is also capable of hydrolyzing both GTP and ATP (4) and has intrinsic kinase activity (5). TGM2 has been implicated in a variety of human diseases including celiac disease, inclusion body myositis, atherosclerosis, and neurodegenerative diseases (6, 7).

References
  1. Gentile, V. et al. (1991) J. Biol. Chem. 266:478.
  2. Chen, J.S.K. and K. Mehta (1999) Internat. J. Biochem. Cell Biol. 31:817.
  3. Griffin, M. et al. (2002) Biochem. J. 368:377.
  4. Lai, T.S. et al. (1998) J. Biol. Chem. 273:1776.
  5. Mishra, S. et al. (2007) J. Biol. Chem. 282:18108.
  6. Kim, S-Y. et al. (2002) Neurochem. Int. 40:85.
  7. Lesort, M. et al. (2000) Prog. Neurobiol. 61:439.
Entrez Gene IDs
7052 (Human); 21817 (Mouse); 56083 (Rat)
Alternate Names
C polypeptide; EC 2.3.2.13; G-ALPHA-h; protein-glutamine gamma-glutamyltransferase 2; protein-glutamine-gamma-glutamyltransferase; TG(C); TG2; TGase C; TGase H; TGase-2; TGase-H; TGC; TGCGNAH; TGM2; Tissue transglutaminase; transglutaminase 2 (C polypeptide, protein-glutamine-gamma-glutamyltransferase); Transglutaminase 2; Transglutaminase C; Transglutaminase H; transglutaminase-2; tTG

Citations for Recombinant Human Transglutaminase 2/TGM2 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. Histone serotonylation is a permissive modification that enhances TFIID binding to H3K4me3
    Authors: LA Farrelly, RE Thompson, S Zhao, AE Lepack, Y Lyu, NV Bhanu, B Zhang, YE Loh, A Ramakrishn, KC Vadodaria, KJ Heard, G Erikson, T Nakadai, RM Bastle, BJ Lukasak, H Zebroski, N Alenina, M Bader, O Berton, RG Roeder, H Molina, FH Gage, L Shen, BA Garcia, H Li, TW Muir, I Maze
    Nature, 2019;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Combined experimental and computational characterization of crosslinked collagen-based hydrogels
    Authors: C Valero, H Amaveda, M Mora, JM García-Azn
    PLoS ONE, 2018;13(4):e0195820.
    Applications: Bioassay
  3. Mechanistic role of transglutaminase-2 in focal adhesions
    Authors: E Png, A Hou, L Tong
    Sci Rep, 2018;8(1):12370.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  4. Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease
    Sci Rep, 2016;6(0):30980.
    Species: Plant - Wheat
    Sample Types: Protein
    Applications: Bioassay
  5. Enzymatically Regulated Peptide Pairing and Catalysis for the Bioanalysis of Extracellular Prometastatic Activities of Functionally Linked Enzymes
    Sci Rep, 2016;6(0):25362.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  6. Secreted osteopontin is highly polymerized in human airways and fragmented in asthmatic airway secretions.
    Authors: Arjomandi M, Frelinger J, Donde A
    PLoS ONE, 2011;6(10):e25678.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay

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