We developed MKN-45 resistant cell line against chemotherapeutic agent and know we would like to assay the activity of the Glutation S Transferase activity in MKN-45 sensitive and resistant cell lines.
How can we do that?
Hello, a lot of commercially available test systems and ready to use kits for detection of Gluthatione S Transferase activity.
https://www.thermofisher.com/order/catalog/product/EIAGSTF
The assay to measure GST activity is relatively simple. You do not need to use kits, if you wish. All you need is a spectrophotometer (able to read at 340 nm) and some cheap reagents, such as phosphate buffer, reduced glutathione and 1-chloro-2,4-dinitrobenzene (CDNB).
See the references below for details:
Welker AF, Moreira DC, Hermes‑Lima M. 2016. Roles of catalase and glutathione peroxidase in the tolerance of a pulmonate gastropod to anoxia and reoxygenation. J Comp Physiol B 186(5):553-568.
Original method reference. Habig WH, Pabst MJ, Jakoby WB. 1974. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 249(22):7130-7139.
Karen A. Darbinyan
thank you dear Dr. karen about your answer to my question.
but we want to assay without using kit. we,re looking for a simple way.
thank you again.
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