Isocitrate Dehydrogenase Assay Services for Drug Discovery
Isocitrate dehydrogenase assays are suited for compound screening and profiling. IDH1 and IDH2 are key metabolic enzymes that catalyze the conversion of isocitrate to a-ketoglutarate (aKG) and cofactor NADPH. Our IDH assays are based on measuring enzyme activity in a coupled system wherein NADPH produced in the initial reaction is a co-factor in the conversion of resazurin to fluorescent resorufin in a secondary reaction.
- Low and large scale as well as high-throughput IDH screening options are available
- Readout: Isocitrate dehydrogenase enzymatic activity
- Custom-tailored IDH assay development possible
- All IDH enzymes are produced in house and available for purchase
List of IDH Assays (11)
| Target | Synonym | Data |
|---|---|---|
| IDH1 G97D | IDH1 G97D | Data sheet |
| IDH1 R100A | IDH1 R100A | Data sheet |
| IDH1 R100Q | IDH1 R100Q | Data sheet |
| IDH1 R132C | IDH1 R132C | Data sheet |
| IDH1 R132H | IDH1 R132H | Data sheet |
| IDH1 WT | IDH1 | Data sheet |
| IDH1 Y139D | IDH1 Y139D | Data sheet |
| IDH2 R140K | IDH2 R140K | Data sheet |
| IDH2 R140Q | IDH2 R140Q | Data sheet |
| IDH2 R172Q | IDH2 R172Q | Data sheet |
| IDH2 WT | IDH2 | Data sheet |
Assay Details
Diaphorase/resazurin-coupled assay
Depending on the mutation status IDH either produces or consumes NADPH. Quantifying the amount of NADPH is therefore a readout for enzyme activity.
For readout, diaphorase utilizes NADPH to convert the dye resazurin to resorufin, a molecule fluorescent at 580 nm.
Setups: Single-dose screening in duplicates or IC50 value determination with 5 or 10 concentrations. Other screening formats are available upon request.
Controls: No inhibitor (DMSO vehicle) control and for every assay, one target-specific control compound is tested in 10-dose IC50 format.
Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.
Report: The raw data, % enzyme activity and control compound IC50 values will be reported in Excel format for single-dose assays. For IC50 orders, raw data, IC50 values, and curve fitting will be delivered in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.
Screening facility: This assay is performed at our screening facility in Malvern, PA, US.
Compound requirements: In brief, for a standard project, 20 µl of a 10 mM DMSO stock or solid material is needed. Less material is needed for large scale screening. Please refer to our FAQs for information regarding compound preparation and shipping.
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