Protocol Online logo
Top : New Forum Archives (2009-): : SDS-PAGE and Western Blotting

Problems with western blot detection - (May/22/2012 )

Hi there,

We have a problem with our western blot detection. Using ecl- film detection the film looks totally black but only the bands appear white (just everything inverted). Does anyone know how this can happen?

Thanks, D


You may be seeing severe over-exposure, either due to using too much antibody, or too strong of an ECL type. What are your antibody dilutions (primary and secondary), and what type of ECL are you using (standard or super-signal/boosters)?