hi, I am working on extracting proteins from pig skin. Initially, I used plain 0.5 M acetic acid solution for extracting proteins and used the solution as such post-centrifugation. but the protein failed to get detected on SDS PAGE. I wish to try the salting out method but am not sure of the steps. can someone please help?
Hi, I'm successfully isolating pig skin proteins using RIPA buffer with protease and phosphatase inhibitors. Also, there can be a problem in the sample preparation before SDS-PAGE.
Vendula Paculová can you point out a few areas where the sample preparation for SDS might have gone wrong...
because I am struggling to troubleshoot the SDS samples
For the SDS I’m using Laemmli buffer and incubating it with protein samples for 10 min at 70 degrees, then centrifugating at 14000g to remove protein agregates. Also 10% separating gel works best for me. What is the protein amount that you are loading?
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