Thioredoxin Reductase BioAssay(TM) Kit (TrxR)

Thioredoxin reductase (TrxR) (EC 1.8.1.9) 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. \n\nThioredoxin Reductase BioAssay(TM) 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 (TNB 2-), which generate a strong yellow color (Lmax = 405 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. \n\nKit Components:\n1. TrxR Assay Buffer: 1x 25ml\n2. TNB Standard: 1x1 vial \n3. DTNB: 1x1 vial \n4. NADPH: 1x1 vial \n5. TrxR Positive Control (~20mU) : 1x1 vial \n6. TrxR Inhibitor: 1x1 vial \n\nStorage and Stability:\nStore kit at -20 degrees C. Protect from light. Allow Assay Buffer to warm to room temperature \nbefore use. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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