Dear all,
I have a bacterial protein that's internalized in human cancer gastric cell (AGS cells). I want to search for its partner so I am planning for a pull down assay.
Which is the best cell lysis buffer do you recommend me to buy ?
And please if you have any tips that would be helpful for the pull down assay I would be grateful.
Thanks in advance,
Mariem
I would recommend the IP Lysis uffer of the company ThermoFiher. This was specifically formulated for this application. It is important to use a lysis buffer without SDS.
If the IP involves only cytoplasmic protein then you can use RIPA buffer with a mild surfactant concentration, put only 0.1-0.5% of Triton X100 and it does the job. For membrane protein you need to add some long chain surfactant with low CMC.
P.S. I will also recommend the IP Lysis by Thermo, it does a neat job.
I may also recommend the Co-IP kit by Thermo which is a one stop job for IP.
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