Isocitrate Dehydrogenase Assay Services

Isocitrate dehydrogenases 1 and 2 (IDH1 and IDH2) are key metabolic enzymes that catalyze the conversion of isocitrate to a-ketoglutarate (aKG) and cofactor NADPH. Reaction Biology provides compound screening against IDH by 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 screening, as well as high-throughput screening options, are available
  • Readout: enzymatic activity
  • Custom-tailored assay development possible
  • All metabolic enzymes are produced in house and available for purchase

List of Targets

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 WT IDH2 Data sheet
IDH2 R172Q IDH2 R172Q Data sheet

Assay Details

Assay Format

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.

Assay setup

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.