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problem with loading the protein on to SDS-Gel


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#1 deep1985

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Posted 18 October 2011 - 10:47 AM

Hi

I am purifying a protein by SP-sepharose cation exchange column. The buffer i use to elute the protein contains potassium chloride in it. Therefore as soon as i add 4X SDS dye to my sample, it precipitates and on heating it becomes sticky and gelatinous. I need to run the protein on the gel to check where my protein is eluted. I cannot load because its too sticky and it wont run on the gel. Any suggestions?

#2 K.B.

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Posted 18 October 2011 - 11:07 AM

1. Can't you use sodium chloride or lithium chloride for elution?
2. You could try buffer exchange with dialysis, ultrafiltration or gel filtration (eg. PD-10 columns).




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