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MAK320 Sigma-Aldrich

D-2-Hydroxyglutarate (D2HG) Assay Kit

sufficient for 200 fluorometric reactions

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Properties

Related Categories Assay Kits by Specialty Area, Cell Biology, Enzymatic Kits for the Quantitation of Nutrients and Metabolites, Metabolomics, Oncology More...
usage   sufficient for 200 reactions (Fluorometric)
shipped in   wet ice
storage temp.   −20°C

Description

General description

The level of D-2-Hydroxyglutarate (D2HG) is low in normal cells and tissues, but is significantly elevated in metabolic diseases, such as the rare autosomal disorder D2HG aciduria. D2HG is mildly elevated in other metabolic disorders including multiple acyl-CoA dehydrogenase deficiency, dihydrolipoyl dehydrogenase deficiency, pyruvate decarboxylase deficiency and pyruvate carboxylase deficiency, various cancers, and in neoplasms with mutations in the isocitrate dehydrogenase 1 (IDH1) and isocitrate dehydrogenase 2 (IDH2) genes. Detection of elevated D2HG is an important biomarker for early diagnosis, prognosis, monitoring and the development of therapeutic strategies against these diseases.

Features and Benefits

• Highly sensitive and rapid enzymatic assay for the detection of D2HG levels in various biological fluids
• Supportive calculator (Click here to download a calculator excel file) Analyze results based on your experimental data!
• Quick instruction bench card - to assure your experimental success
• More experiments in one kit - contains sufficient reagents for 200 tests
• Detection time: only 30-60 minutes

Suitability

Suitable for the detection of D2HG levels in cells and tissues lysates, serum, urine, cultured cells and culture supernatants.

Principle

Developed in partnership with the German Cancer Research Center (DKFZ), the D2HG Assay Kit is a rapid and sensitive enzymatic assay for the detection of D2HG levels in various biological fluids: serum, urine, cell culture supernatants, and cell or tissue lysates. The assay, originally developed by Balss et al., 8 is based on the oxidation of D2HG to α-ketoglutarate (αKG) by the enzyme (D)-2-hydroxyglutarate dehydrogenase (HGDH) coupled to the reduction of NAD+ to NADH (see Figure 1). The amount of NADH formed is then quantitated by the diaphorase mediated reduction of resazurin to the fluorescent dye resorufin (λex = 540 nm/λem = 590 nm).

Legal Information

This product is sold under license from the German Cancer Research Center (DKFZ) and University Clinic of Heidelberg. Use of this product is covered by certain US and foreign patents, including US Pat. No. 9,487,815, EP Pat. No. 2 820 145 and foreign equivalents. Use of this product is for research purposes only.

Kit component only

Description

Product #

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D2HG Standard, 10mM aqueous solution    
D2HG Reaction Mixture, For preparation of 7.5 ml reaction mixture    
Resazurin Solution, 125 μM solution in 50% DMSO    
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
Protocols & Articles
Peer-Reviewed Papers
15

References

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