In my studies I want to evaluate heterodimer formation of different STAT molecules after stimulation with different cytokines. What buffer can I use since those proteins are phosphorylated and the dimer interaction (non-covalent) needs to remain intact for the Co-IP.
Can you provide recipes for buffers that you have used for similar purposes? Thank you!
Hello Nils Ott
You need to prepare the lysis buffer such that it will minimize protein denaturation and at the same time does not affect the target and the associated protein. Please refer to the link below for more information on how to go about with the lysis buffer preparation for Co-IP. You need to add protease inhibitors as well as phosphatase inhibitors in the buffer.
https://www.abcam.com/protocols/immunoprecipitation-protocol-1
Best Wishes.
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