unspecific binding
 

Hello.
Can anyone tell me if there are sequences that can bind unspecific to Protein A Agarose more easy then others? Or is it just a matter of size?
Thank you very much.

Re: unspecific binding
 

Hello,
there may be some sequences that do it more often that others. Not sure about size.

Have you tried non-specific competitors? Using BSA? other methods to block non-specific binding such as harsher conditions?

cheers

Re: unspecific binding
 

My Protein A Agarose is pre-blocked with salmon sperm DNA, and before I add the immunoprecipitating AB, I pre-clear the eluates with Protein A Agarose, that I discard afterwards.

Re: unspecific binding
 

hmm any progress on this I may also conduct Chromatin Immunoprecipitation soon... maybe has to do with the Protein A?

Re: unspecific binding
 

Did you check the sheared DNA on gel? what's the size distribution?

Also,You generally have much less of these bacgkround problems if you use magnetic beads instead of agarose.

Re: unspecific binding
 

Did you check the sheared DNA on gel? what's the size distribution?

Also,You generally have much less of these bacgkround problems if you use magnetic beads instead of agarose.