Negative stain directly after Ni NTA?

04 April 2018 1 6K Report

Hi,

I am working on a membrane protein (of around 153kDa) which I am eluting by 300mM imidazole (Elution volume-2 ml). I loaded 20ul of my elution fraction on SDS PAGE and I could see my protein purity upto 80 percent (before SEC). Can I make negative staining grids directly? In other words, will imidazole affect the negative stain?

Ke-Wei Zhao

Haaris: I don't see a reason for not making direct negative staining, as imidazole can easily wash out gel in solution.

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Science method

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