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purifying HA-tagged membrane protein (receptor)

purifying HA-tagged membrane protein (receptor) - Protein Science

purifying HA-tagged membrane protein (receptor) - Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.


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Old 01-26-2009, 08:14 PM
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Default purifying HA-tagged membrane protein (receptor)



Hello all,

I have HA-tagged a Her2 construct that I made, and expressed it in 293t cells and have a total protein lysate now. My HA tag is in between the Her 2 protein and a red flourescent tag, I was wondering if it is ok to go ahead and denature the lysate before I use an anti-HA tag affinity matrix to pull out the protein of interest? I want to make sure the protein is linearized so that the HA tag won't be occluded to the antibody in the matrix. I am going to run the purified eluate on a gel and send the band for N-ter sequencing.

Thanks!
Toby
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