Transglutaminase 2/TGM2 Assay Kit (Colorimetric) Summary
The Transglutaminase 2/TGM2 Assay Kit uses biotinylated T26 peptide (Biotin-pepT26) as the first substrate (amine-acceptor/acyl-donor) and an amine-donor/acyl-acceptor as a second substrate.
Samples suspected of containing TG2 are incubated with calcium, dithiothreitol (DTT) and Biotin-pepT26 in the wells of microtiter plates containing the amine substrate. In the presence of TG2, the gamma carboxamide of the glutaminyl residue of the biotin-pepT26 is incorporated into the amine substrate to form biotinylated isopetide bound.
The system is coupled to Streptavidin labelled peroxidase (SAv-HRP). SAv-HRP is revealed using H2O2 as HRP substrate and tetramethyl benzidine as electron acceptor (chromogen).
Measurement of tissue Transglutaminase (tTG/TG2) with low or no interference with other TGs isoforms.
Assay Kit (Colorimetric)
Packaging, Storage & Formulations
Storage of components varies. See protocol for specific instructions.
- Reaction Buffer (Biotin-pepT26/CaCl2) (lyophilized)
- Opr0036 : Recombinant human TG2 (positive control) (lyophilzed)
- Wash Buffer 10X (30 ml)
- Blocking Reagent (12 ml)
- HRP Substrate (12 ml)
- Microtiter strips coated with amine substrate (12x8 strip microtiter wells)
- Enzyme Tracer (SAv-HRP) (50 ul)
- Diluent Buffer 10X (10 ml)
- DTT (0.2 ml)
- EDTA (Negative Control) (1 ml)
The kit is shipped on ice. Upon arrival, the DTT (R1), the Reaction Buffer (R3) and Enzyme Tracer (R4) should be stored at -20C. All the other components of the kit should be kept at 4C. When stored properly, these stock solutions are stable for at least 24 months. OTHER SUPPLIES REQUIRED: - Recombinant human TG2 - TG2 inhibitor. This is recommended as negative control for screening of TG2 inhibitors.
Alternate Names for Transglutaminase 2/TGM2 Assay Kit (Colorimetric)
- C polypeptide
- EC 188.8.131.52
- protein-glutamine gamma-glutamyltransferase 2
- TGase C
- TGase H
- Tissue transglutaminase
- transglutaminase 2 (C polypeptide, protein-glutamine-gamma-glutamyltransferase)
- Transglutaminase 2
- Transglutaminase C
- Transglutaminase H
Transglutaminases (EC. 2.3.213, R- glutamyl-peptide amine g-glutamyl-transferase) are a family of calcium dependent enzymes which catalyse an acyl transferase reaction between the gamma-carboxamide group of peptide bound glutamine and various primary amines. In mammals, at least eight active transglutaminase (TGs) isoenzymes have been described so far. They are widely distributed in various organs, tissues and body fluids. Among them, transglutaminase type 2 (TG2) is distinguished from others TGs by its functional versatility and ubiquitous expression pattern in mammalian tissues. This isoenzyme is involved in a variety of roles including stabilization of intra and extracellular matrices and crosslinking of cell envelopes in apoptosis. It has also been associated with a large number of pathological conditions such as fibrosis, celiac disease, neurodegenerative disorders, inflammatory processes in sepsis, and in carcinogenesis of hepatocellular and ovarian carcinoma. This consideration has led to the development of several tests for either research or clinical purpose. Among the various strategies developed to measure TG activity, the most sensitive and accurate quantification methods, although non-discriminatory, have been the radioactive and fluorimetric assays. Those methods detect the incorporation of labeled molecules, such as [3H/14C]-putrescine or monodansyl-cadaverine respectively, into glutamyl substrates such as casein or synthetic peptides. Solid phase assays developed so far in similar principles have been compromise by a high background signal, low sensitivity compare to radiolabeling methods and the lack of the specificity. This test kit overcomes these problems and constitutes a highly sensitive and specific TG2 solid-phase microassay. The TG2-Covtest uses a biotinylated preferred first substrate of TG2 (biotin-pepT26) as amine-acceptor, and spermine as second substrate (amine-donor) of the enzyme. Samples suspected of containing TG2 are incubated with calcium, dithiothreitol (DTT) and biotin-pepT26 in the wells of microtiter plates to which spermine has been covalently coupled. In the presence of TG2, spermine is incorporated into the g carboxamide of the glutaminyl residue of biotin-pepT26 to form a biotin-pepT26- g-glutamyl spermine. Enzymatic reaction is determined by its interaction with Streptavidin labelled peroxidase (SAv-HRP). Following a wash step to remove any unbound enzyme reagent, a substrate solution for SAv-HRP containing H2O2 and tetramethyl benzidine as electron acceptor (chromogen) is added and color developed. The color intensity is directly proportional to the TG2 activity in the sample.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed
for 6 months from date of receipt.
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Publications for Transglutaminase 2/TGM2 Kit (NBP1-37008)(3)
Showing Publications 1 - 3
|Publications using NBP1-37008
| Jung HJ, Chen Z, Wang M et al. Calcium blockers decrease the bortezomib resistance in mantle cell lymphoma via manipulation of tissue transglutaminase activities Blood 2012 Mar 15 [PMID: 22294726]
| Perez Alea M, Kitamura M, Martin G, Thomas V, Hitomi K, El Alaoui S. Development of an isoenzyme-specific colorimetric assay for tissue transglutaminase 2 cross-linking activity. Anal Biochem. 389(2):150-6. 2009 Jun 15. [PMID: 19318081]
| Thomas V, El Alaoui S, Massignon D, Clement S, Simonet F, Quash G. Development and evaluation of a modified colorimetric solid-phase microassay for measuring the activity of cellular and plasma (Factor XIII) transglutaminases. Biotechnol Appl Biochem. 43(Pt 3):171-9. 2006 Mar. [PMID: 16313237]
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