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Transglutaminase Activity Assay Kit (Colorimetric)

Validated using many sample types

WARNING: This product can expose you to chemicals including Trichloroacetic Acid, which is [are] known to the State of California to cause cancer.  For more information go to
Catalog #: K571

Product Details

Cat # +Size K571-100
Size 100 assays
Detection Method Absorbance (525 nm)
Applications Measurement of Transglutaminase activity
Features & Benefits • Simple, rapid & convenient
• The assay can detect transglutaminase activity ~10 µU or 80 ng of recombinant hTG2 enzyme
Kit Components • TG Assay Buffer
• Homogenization Buffer (10x)
• 1M DTT
• Donor Substrate
• Acceptor Substrate
• Hydroxamate Standard
• Stop Solution
• Positive Control
• Plate Sealer
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Transglutaminases (EC are calcium dependent enzymes that catalyze the post-translational modification of proteins by formation of isopeptide bonds. This occurs either through protein cross-linking via formation of γ-glutamyl-ε-lysine bonds or through incorporation of primary amines at selected peptide-bound glutamine residues. The transglutaminase enzyme family comprises the intracellular forms (TG1, TG3 and TG5) expressed mostly in the epithelial tissue; TG2 which is both intracellular and extracellular and expressed in various tissue types; TG4 which is expressed in the prostate gland; factor XIII which is expressed in blood; TG6 and TG7, whose tissue distribution is unknown and band 4.2 (lacking enzymatic activity) which is present on erythrocyte membranes. Transglutaminases also exhibit GTPase, phosphodiesterase and protein kinase activity. Transglutaminases are associated with certain neurological and autoimmune disorders and also cancer. BioVision’s Transglutaminase Activity Assay kit utilizes the deamidation reaction of the transglutaminase enzyme with a donor and acceptor substrate resulting in the formation of a hydroxamate product. The hydroxamate product reacts with the Stop Solution forming a purple complex that can be measured colorimetrically at 525 nm. The limit of quantification of this assay is ~10 µU or 80 ng of recombinant hTG2 enzyme.

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What is the nature of the donor and acceptor molecules? What is the overall principle of this assay?
Exact names of the donor and acceptor substrate are proprietary to BioVision and hence cannot be disclosed. However, we use an amine acceptor substrate and amine donor substrate. In the presence of transglutaminase, the donor amine is incorporated into the acceptor amine to form a product, which develops a colored complex with iron, detectable at 525 nm.
Is the chloride reagent supposed to come in bright purple color? or should it be rather colorless?
The chloride reagent is bluish in colour to begin with. The final colour developed is going to be intense blue which can be detected at 620 nm.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration?
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.