Is it possible to correct an impure protein sample that was already diluted in 2x laemmli buffer ?

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Gautam Kumar Ginjupalli

Dear Steingrimur,

I have protein extracted from fat samples of fat. I am finding smeared bands on my western blots. Forums on research gate had few tips. But, my protein was already diluted in the laemlli buffer. I would like to know if it is possible to desslt my samples and get rid of any residual fats that might be remaining in the s samples.

Thanks in advance for your time and concern.

Steingrimur Stefansson

Hi Gautam, to begin with, desalting columns will not get rid of the lipids in your samples.

I would recommend to treat your sample with an equal volume of cold methanol (at -20oC).

The proteins will precipitate, but the lipids will stay in the methanol phase.

Dry the precipitate and run on a gel.