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Pool down assay

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#1 Vijaytailor

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Posted 24 March 2015 - 09:05 PM

Hi there,

I am performing a pool down assay using ProteinS sepharose,  in that i am getting my protein of intereast, and Anti body also in the elute.

My problem is that i want to elute my protein but not the Antibody in it. Can any one suggest any protocol for this?



#2 mdfenko

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Posted 25 March 2015 - 05:27 AM

you can crosslink the antibody to protein a sepharose (i've used protein a, you can also use protein g, never heard of protein s sepharose) prior to binding your protein of interest then elute with either low pH or high pH (whichever works better for you). the antibody will remain attached to the immobilized protein.


Edited by mdfenko, 25 March 2015 - 05:27 AM.

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