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Abcam's Thioredoxin Reductase Assay Kit provides a convenient colorimetric assay for detecting TrxR activity in various samples. In the assay TrxR catalyzes the reduction of 5, 5'-dithiobis (2-nitrobenzoic) acid (DTNB) with NADPH to 5-thio-2-nitrobenzoic acid (TNB2-), which generate a strong yellow color (λmax = 412 nm). Since in crude biological samples other enzymes, such as glutathione reductase and glutathione peroxidase, can also reduce DTNB, therefore, TrxR specific inhibitor is utilized to determine TrxR specific activity. Two assays are performed: the first measurement is of the total DTNB reduction by the sample, and the second one is the DTNB reduction by the sample in the presence of the TrxR specific inhibitor. The difference between the two results is the DTNB reduction by TrxR.
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Thioredoxin reductase (TrxR) (EC 18.104.22.168) is a ubiquitous enzyme which is involved in many cellular processes such as cell growth, p53 activity, and protection against oxidation stress, etc. The mammalian TrxR reduces thioredoxins as well as non-disulfide substrates such as selenite, lipoic acids, lipid hydroperoxides, and hydrogen peroxide.
|TNB Standard||Brown||1 vial|
|TrxR Assay Buffer||WM||1 x 25ml|
|TrxR Inhibitor||Clear||1 vial|
|TrxR Positive Control||Green||1 vial|
Our Abpromise guarantee covers the use of ab83463 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|
TNB (Thionitrobenzoic acid) measured in mouse tissue lysates after 25 min. incubation showing quantity (mU) per mg protein of tested sample.
TNB (Thionitrobenzoic acid) measured in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample
TNB (Thionitrobenzoic acid) measured in cell lysates after 25 min. showing quantity (mU) per 1 mln of tested cells
Standard curve (colourimetric) : mean of duplicates (+/-SD) with background readings substracted
TNB (Thionitrobenzoic acid) measured in cell lysates showing quantity (nmol) per 1 mln of tested cells
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