Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum

Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum (http://www.molecularstation.com/forum/)
-   ChIP Chromatin Immunoprecipitation Forum (http://www.molecularstation.com/forum/chip-chromatin-immunoprecipitation-forum/)
-   -   unspecific binding (http://www.molecularstation.com/forum/chip-chromatin-immunoprecipitation-forum/47531-unspecific-binding.html)

Chip 11-26-2008 12:27 PM

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.

admin 11-26-2008 08:05 PM

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

Chip 11-27-2008 11:45 AM

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.

moleculardude 12-03-2008 03:05 PM

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

Joegreen 05-08-2009 09:51 AM

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.

Joegreen 05-08-2009 09:52 AM

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.


All times are GMT. The time now is 12:02 AM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved

Page generated in 0.09195 seconds with 11 queries