unspecific binding

[Chip]

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.

[admin]

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

[Chip]

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.

[moleculardude]

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

[Joegreen]

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.

[Joegreen]

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.