Hi everyone. I tried concentrating and precipitating my protein using ProteoExtract Protein Precipitation kit. I did not have any of the buffers suggested to dissolve so I used RIPA buffer. Unfortunately the protein pellet did not dissolve. Any suggestions of how to rescue my protein? Need to eventually check protein concentration and use for western blot. Thanks!
Use 0.05 M Tris buffer set to pH 7.5 at 3-40 degree Celcius. Hope it shall work. For SDS page directly add sample buffer to the protein, heat for 3 min in boiling water bath and load on the gel.
Could you dissolve the precipitated protein? You can warm the buffer to 37 C, which will increase the solubility of a protein.
The responses above are useful. In addition, do you have access to a bath sonicator. Gentle sonication can assist with solubilising precipitated peptides and protein.
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