relaxin wrote:Are your Markers pre-stained? Your peptide is only 1 kDa, it may run out of the gel or may not bind to PVDF membrane. Coomassie blue stain will not work. You need reversible MemCode™ Stain from Pierce.
You may try to probe with antibody against a reference protein (beta-actin) first. If it works, then probe with your precious antibody.
mdfenko wrote:i used to separate enkephalins with tris-tricine gels.
0.2um pore is okay. you can get smaller pore but that shouldn't be necessary. i used semi-dry transfer for 15-20 minutes at the recommended current density (mA/cm2) for the apparatus.
kavizz wrote:Can you please share your protocol in detail. I am trying to detect a 7kDa protein and am having problems with it.
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