Smeared Bands after Ponceau Staining? - (Jul/13/2013 )

Hello,

I have observed smeared looking bands on my Ponceau-stained PVDF after transfer. My samples are from bone, so I'm not sure what is the cause for this and/or if I should be concearned. Other than ~3-4 defined bands, the rest just looks smeared... (please see photo)?

I'm not sure if this is what protein degradation looks like, but I do add appropriate protease/ phosphatase inhibitors to my buffer?

Any ideas/ suggestions would be greatly appreciated!

Thanks all,

Beth

It is quite likely to just be a level of protein thing, you could test by coomassie staining a gel and see if you get the same pattern.

it could also be due to degraded and/or aggregated proteins.

do you boil the samples in the sds loading buffer? if so, try incubating at 60-70C for 10-20 minutes instead of boiling.

Thank You I'll try this today!