The IDH1(R132H) Assay Kit comes in a convenient 96-well format, with enough purified recombinant IDH1(R132H) enzyme, α-KG, NADPH, NADPH detection reagents, and assay buffer for 100 enzyme reactions.
Materials Required But Not Supplied
NADPH (optional)* Microplate reader capable of reading Fluorescence Adjustable micropipettor and sterile tips
*Note: NADPH is not stable for multiple freeze/thaw cycles. We have provided the reagent as 2 x 100 μl to allow you to use only half the plate at a time. If you plan to use the same plate for additional experiments, please purchase additional NADPH and prepare a fresh 500 μM stock in H2O at the time of the assay.
Isocitrate dehydrogenases are enzymes that catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. Mutations in isocitrate dehydrogenases 1 and 2 (IDH1 and IDH2) have been reported in a multitude of human cancers. While the wild type IDH1, produces α-ketoglutarate (α-KG) through the reduction of NADP+ to NADPH, the IDH1(R132H) mutant catalyzes conversion of α-ketoglutarate to 2-hydroxy-glutarate (2-HG) by oxidizing NADPH to NADP+. Genomic studies have identified the IDH1(R132H) mutation in various cancer types, including glioma, chondrosarcoma, and AML as well as a small percentage of prostate, lung and colon cancers.
References
Wang, F. et. al. Science340:622-626 (2013).
Storage/Stability
Stable at least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Instructions for Use
See assay kit data sheet for detailed protocol.
Shipping Temperature
-80°C
Warnings
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.