Hi,
After using a stripping buffer for 12 min at rtp, my band of interest (~50kDa) still appeared together with the housekeeping band (Beta actin). Is this still ok? Or is there a way to remove it altogether?
Use the strong stripping condition with Tris;beta mercapto ethanol and SDS incubate in 50'C/10 mins.
Other concerns check you secondary antibody if both are rise from same species will inefficient striping will give multiple bands.
Check the blot after stripping (Incubate with ECL and check for signal if no signal proceed for another antibody)
If no other ways is there try to cut membrane and leave only the portion of your interest.
stripping was never complete in my hands for strongly expressed proteins. do not forget that the stronger the stripping, the more of the blotted proteins will be washed away along with the antibodies.
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