Hi,
I'm considering using polyacrylamide gel percentages as a tool to remove higher molecular weight proteins for my Ph.D. project.
In reference, polyacrylamide % helps resolve proteins of interest based on molecular weight: 12.5% for 10-70 kDa proteins, 10% for 15-100 kDa proteins, etc. For examples, please see the table @ https://www.abcam.com/protocols/electrophoresis-for-western-blot Or https://www.novusbio.com/support/support-by-application/western-blot-sds-page.
With this information, I ran the samples on 4 different percentage gels - 12%, 13.5%, 14%, and 15%. Below are related images for your perusal. Unfortunately, despite there being a good separation, the higher percentage gel is not cutting off the higher molecular weight proteins. Almost all gels show proteins including those above 70 kDa.
In this regard, how do I cut off higher molecular weight proteins using polyacrylamide gels percentage for SDS PAGE?
I greatly value your feedback, suggestions, and consideration.
Thank you very much for your time, in advance.
Raja
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