Due to the high cost of Magnetic IP Kits, our group has decided to make the buffers ourselves. We intend to use SDS Lysis buffer for cell lysis, which should work just fine. However I struggle to find clear information on which wash buffer to use. There are lots of suggestions includung plain PBS, PBS-T, Lysis buffer or a self-made wash buffer (Tris, EDTA, EGTA, Triton x100, NaCl and protease inhibitors) Does anybody have experience or suggestions for which wash buffer to use?
Thank you in advance
I assume you're using protA/G mag beads. Pretty robust, but depends on your IP Ab. Start by using some anionic detergent (Triton X100, or NP40 around 0.1% but could be higher) and isotonic conditions, especially if you intend to move to Co-IP xpts. You should include protease inhibitors. Rest is likely optional.
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