Question (74378) | Alpha Ketoglutarate (alpha KG) Assay Kit (ab83431)

Go to datasheet (ab83431)

Question

What is the suggested way to lyse cells for use with this kit but not interfere with enzymatic activity?

Answer

The most efficient results will be had if the homogenate is made with the assay buffer provided with the kit. Rapidly homogenize ~2 x 10^6 cells with 100 µl of ice cold α-KG Assay Buffer using a dounce homogenizer. Deproteinate the samples so you can get rid of any interfering proteins including the enzymes. This is a small molecule analyte assay and cellular enzymes can interfere with the signal development

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