Dear expert peers,
which is the best lysis buffer to lyse the isolated platelet pellets. We were using RIPA buffer in earlier analyses (150 mM sodium chloride, 25 mM Tris-HCl pH 7.6, 1% Tergitol-type NP-40, 0.1% sodium dodecyl sulfate, 1% sodium deoxycholate).
Now, looking for a readymade better alternative.
Any suggestions with the product name or recipe would be highly appreciated.
DISCLAIMER: I don't work with platelets, so this may not be better for what you are doing specifically.
However, I have been working on optimizing our cell lysis protocol lately. I have found with the buffers that I have looked at, that M-PER with a protease inhibitor pill has done the best. I have also looked at adding 100 mM KCl to the M-PER PPi solution but it's not clear yet if that definitively helps yet or not (I haven't tried to quantitate it and the bands are similar in size).
Just so you know the cells I am doing this in is a zebra fish cell line, and I am looking at eIF4e5 with anti-hypusine antibody as my primary on a western and all lanes are loaded with the same number of cell equivalents. If you have any questions feel free to let me know, I hope this helps!
Dear Dr. Ali and Kaila. Thank you very much for your valuable suggestions. Much appreciate.
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