The PROTEOSTAT® Thioredoxin-1 assay kit provides a simple, homogenous assay for screening modulators of Trx-1 enzymatic activity in microplates. This is accomplished by monitoring the Trx-1 catalyzed reduction of insulin in the presence of Dithiothreitol (DTT), resulting in the formation of insulin aggregates which then bind avidly to the red-emitting fluorogenic PROTEOSTAT® Trx-1 detection dye. The fluorescence-based assay provides a vastly improved assay signal window, improved lower detection limit, and superior Z’-score (>0.8) relative to turbidity measurements. Intra-plate and inter-plate CVs using this assay are typically 3-6%. Human recombinant thioredoxin-1 enzyme and the thioredoxin inhibitor, aurothiomalate, are included in the kit, as well as all reagents necessary for monitoring changes in Trx-1 activity. The PROTEOSTAT® Thioredoxin-1 assay kit is capable of providing a quantitative readout of Trx-1 enzymatic activity in a robust and high-throughput fashion and can be applied to identification of Trx-1 inhibitors from chemical libraries.
Product Details
| Applications: | Fluorescent detection
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| Application Notes: | This kit provides a simple, homogenous assay for screening modulators of Thioredoxin-1 enzymatic activity. |
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| Quality Control: | Using the procedure described in the manual, a sample from each lot of PROTEOSTAT® Thioredoxin-1 assay kit is used to assay the following: (1) Insulin with Trx-1; (2) Insulin without Trx-1; and (3) Insulin with Trx-1 and Aurothiomalate. The Z’-factor is greater than 0.8. Trx-1 is inhibited by at least 50% with Aurothiomalate. |
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| Use/Stability: | Upon receipt, store the Thioredoxin-1 at -80°C. Store the remaing reagents at ≤-20°C protected from light. |
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| Handling: | Protect from light. Avoid freeze/thaw cycles. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Contents: | PROTEOSTAT® PDI detection reagent: 20 μL
Thioredoxin-1 (human) (recombinant): 2 vials x 80 μL
Insulin: 2 vials x 1.8 μmol
Aurothiomalate (inhibitor): 2 vials x 25 nmol
PBE buffer: 25 mL
Stop reagent: 1 mL
DTT: 2 vials x 1.3 mL
Deionized water: 5 mL |
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| UniProt ID: | P10599 |
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
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Application(s): RAW264.7 and THP-1 cells,
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Thioredoxin o-mediated reduction of mitochondrial alternative oxidase in the thermogenic skunk cabbage Symplocarpus renifolius: Y. Umekawa & K. Ito; J. Biochem.
165, 57 (2019),
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Full Text
Reactive oxygen species, glutathione, and thioredoxin influence suberoyl bishydroxamic acid-induced apoptosis in A549 lung cancer cells: B.R. You, et al.; Tumour Biol.
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Abstract;
PX-12 inhibits the growth of A549 lung cancer cells via G2/M phase arrest and ROS-dependent apoptosis: B.R. You, et al.; Int. J. Oncol.
44, 301 (2014),
Application(s): Measurement of Trx-1 activity,
Abstract;
