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Co-IP problems with band sizes


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#1 malena.a

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Posted 30 May 2009 - 07:05 AM

Hey
I would like to get some opinions on my problem. I have performed standard co-immunoprecipitation with the antibody against protein A. I want to confirm previously published protein protein interactions. Between the above mentioned protein A and "my protein B'. However the input sample with antibody anti-B on WB gives band size 90 kda which is correct, but using the same antibody after Co-IP I see the band about 75-80 kDa. Is it possible that in complexes the protein got smaller? or is it more likely unspecific band. Thank you for time and efforts.

#2 mikew

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Posted 01 June 2009 - 02:03 PM

Photo?
Also, the salt concentration and detergents in your samples can affect running mobility.
Try to ensure that buffers in input and IP are consistent.




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