Transglutaminase 3 (TG3), also known as epidermal Transglutaminase (Tgase E), belongs to the family of Transglutaminase enzymes that catalyze the posttranslational modification of proteins via calcium dependent cross-linking reactions (1-3). TG3 is involved in the formation of the cornified envelope in skin keratinocytes (4). It functions to cross-link structural proteins during epidermal terminal differentiation. TG3 has been implicated as the dominant autoantigen in dermatitis herpetiformis (5). TG3 activation requires proteolysis of the 77 kDa zymogen into two fragments of approximately 50 and 27 kDa to form the active enzyme (1).
Recombinant Human Transglutaminase 3/TGM3 Protein, CF
R&D Systems | Catalog # 4604-TG
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Key Product Details
- R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Transglutaminase 3/TGM3 Protein (4604-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
Applications
Enzyme Activity
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Product Specifications
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Transglutaminase 3/TGM3 protein
Ala2-Glu693 (Gly654Arg), with an N-terminal Met and 6-His tag
Ala2-Glu693 (Gly654Arg), with an N-terminal Met and 6-His tag
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
No result obtained
Predicted Molecular Mass
72 kDa
SDS-PAGE
72 kDa, reducing conditions
Activity
Measured by its ability to cleave a synthetic peptide Benzyloxycarbonyl-Gln-Gly and NH2OH.
The specific activity is >450 pmol/min/µg, as measured under the described conditions.
The specific activity is >450 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
4604-TG
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol. |
| 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.
|
Background: Transglutaminase 3/TGM3
References
- Kim, I.G. et al. (1993) J. Biol. Chem. 268:12682.
- Griffin, M. et al. (2002) Biochem. J. 368:377.
- Lorand, L. and R.M. Graham (2003) Nat. Rev. Mol. Cell Biol. 4:140.
- Eckert, R.L. et al. (2005) J. Invest. Dermatol. 124:481.
- Sardy, M. et al. (2002) J. Exp. Med. 195:747.
Alternate Names
TG3, TgaseE, TGM3
Gene Symbol
TGM3
UniProt
Additional Transglutaminase 3/TGM3 Products
Product Documents for Recombinant Human Transglutaminase 3/TGM3 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Transglutaminase 3/TGM3 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Transglutaminase 3/TGM3 Protein, CF
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Protocols
View specific protocols for Recombinant Human Transglutaminase 3/TGM3 Protein, CF (4604-TG):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 0.15 M NaCl, 0.05% Brij, pH 7.5 (TCNB)
- Assay Diluent: Deionized water
- Recombinant Human Transglutaminase 3/TGM3 (rhTGM3) (Catalog # 4604-TG)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- Substrate: Z-Gln-Gly (Sigma, Catalog # C6154), dissolve 500 mM in deionized water, then adjust to pH 9.0 with NaOH
- MES, pH 6.0, 400 mM stock in deionized water
- DTT (Sigma, Catalog # D-0632), 200 mM stock in deionized water
- CaCl2, 200 mM stock in deionized water
- 1 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
- Combine the following volumes per reaction vial: 10 µL of 500 mM Substrate, 50 µL of 400 mM MES, pH 6.0, 5 µL of 200 mM DTT, 5 µL of 200 mM CaCl2, and 10 µL of 1.0 M Hydroxylamine Hydrochloride right before running assays. 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.
- Dilute rhTGM3 to 0.2 mg/mL with 2 ng/µL Thermolysin in Assay Buffer.
- Incubate 30 minutes at room temperature.
- Dilute rhTGM3 to 0.05 mg/mL in Assay Diluent.
- Mix 20 µL of the diluted rhTGM3 with 80 µL Substrate Mixture. For the Blank mix 20 µL of Assay Diluent with 80 µL Substrate Mixture.
- Incubate at 37 °C for 2 hours.
- After incubation, stop the reaction with 400 µL of the Stop Solution. Mix well.
- Centrifuge at top speed for 2 minutes in a microcentrifuge.
- Load 200 µL of the supernatant into a plate.
- Read at 525 nm (absorbance) in endpoint mode.
- 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 g-monohydroxamate (Sigma, Catalog # G2253).
Per Well:
- rhTGM3: 0.4 µg
- Substrate: 10 mM
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