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

Blotchy Blots - (Apr/12/2016 )



I'm wondering does anyone know what would be causing these "blotchy" blots. Semi dry transfer, all new reagents, new buffer etc. Transferred successfully as rainbow markers all transferred. You can see in the images when I ponceau stain there is this white patch in the middle. This is happening all the time. When I develop the blots there is always a high background/they are dirty.


Any ideas?



Attached Image

Attached Image


a few possible reasons:


poor contact in that area during transfer


shaking during staining/destaining is insufficient, allowing membrane to stick to staining vessel


pouring destain solution (water when using ponceau) directly onto blot at that spot


many thanks mdfenko.