Hello everyone,
I've been experiencing a problem with my western blots recently. For some reason my PVDF membrane is extremely blurry after imaging with barely of my bands showing up. I've been following the exact same protocol (BioRad) each time and never had this issue with my previous blots. Can anybody provide any advice in troubleshooting this?
What is your BioRad protocol development/ imaging? Did the camera on the BioRad unit get changed or filters added/removed? The image looks over exposed/ gain too high, my images I can see the membrane centered in a black field not a white one like yours. I would suggest eliminating hardware problems 1st, then adjust your development/ imaging protocol only if necessary.
Hi Helen,
it seems that you have problem with the image processing. or like Jeffrey has mentioned, your image seems to be overexposed. Did you see the band formed on the PVDF membrane with naked eye?
I think you should be able to capture the membrane directly using a good resolution camera. having said that, we have used a normal image scanner (the one for the document) to scan most of our gels and membrane and they came out pretty good. you can try scanning using different setting: black/white or colour. Good Luck!
Hi Helen, you can do a Poncau-S staining of the blot prior to the imaging to see if the problem is the transfer or the imaging.
- Badges
- Science method