white patch in TLC with immunostaining

Patricia Román-Carrasco @Patricia-Roman-Carrasco

27 July 2017 1 6K Report

Hello!

Quite often, when I ve done a TLC with immunostaining (I overlay with poly isobutyl methacrylate), I get in the picture those ugly huge bright/white areas instead a clean picture!

Does someone know why they do appear and how can I avoid it?

Thanks a lot!

Thoms Payne

I am not familiar with TLC with immunostaining, however, it looks as though your initial blots were not fully dry. Also if those chemicals separated out by TLC are not somehow fixed to the TLC plate, then submerging the TLC plate in another solvent will cause the band to diffuse (broadened) if not dried fast enough. Again, I am not sure of your process and so it is hard to pinpoint what went awry.

https://www.youtube.com/watch?v=bweDL6BgvAo

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