Thioredoxin Reductase Bacteria Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Mammalian thioredoxin family members include thioredoxin-1 (Trx1), mitochondrial thioredoxin-2 (Trx2), and a larger thioredoxin-like protein, p32TrxL. Thioredoxin is reduced by NADPH and thioredoxin reductase and, in turn reduces oxidized cysteine groups on proteins. When thioredoxin levels are elevated, there is increased cell growth and resistance to the normal mechanism of programmed cell death. Thioredoxin reductase (TrxR, EC 1.6.4.5) is a family of glutathione reductase-like homodimer selenol-containing flavonenzyme. The redox activity of this catalytic site is necessary for the biological activity of thioredoxin. An inhibitor of thioredoxin reductase may be useful in the treatment of cancer.
Family:
Oxidoreductase
Sub Family:
Reductase
Class:
Bacterial
Organism:
Bacteria
Construct Details:
Endogenous
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrophotometry
Measured Response:
Absorbance
Testing Information
Procedure Summary:
Bacterial (E. coli) thioredoxin reductase is used. Test compound and/or vehicle is preincubated with 190 nM enzyme‡ in phosphate buffer pH 7.4 for 15 minutes at 25⁰C. The reaction is initiated by addition of 1 mM 5,5'-Dithiobis(2-nitrobenzoic acid) for another 60 minute incubation period. Determination of the amount of 5-mercapto-2-nitrobenzoic acid formed is read spectrophotometrically at 405 nm. Compounds are screened at 10 μM.
Substrate:
1000 μM 5,5'-Dithiobis(2-nitrobenzoic acid) (DTNB)
Incubation:
60 min at 25°C
Control Inhibitor:
AURANOFIN
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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