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SDS-PAGE Gel Electrophoresis Forum SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


SDS-PAGE of Secreted Proteins

SDS-PAGE of Secreted Proteins - SDS-PAGE Gel Electrophoresis Forum

SDS-PAGE of Secreted Proteins - SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


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Old 01-13-2008, 11:12 AM
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Exclamation SDS-PAGE of Secreted Proteins



HIYA!!

IM bak for a question.

I am studying secreted proteins now. My weird question is if I isolate secreted proteins from media, will I have enough of them to run on an sds-page?
I did protein concentration and it seems like there arent many proteins after washing and treating.

I grow the cells for a few hours, day, and days and still low protein.


Also what is the best method to visualize low abundance proteins? Western blot? Coomasie? SYPRO? Silver stain?

thnx guys.
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Old 01-13-2008, 05:06 PM
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Default Re: SDS-PAGE of Secreted Proteins

well
if enough quantity of protein is secreted in the media then you can visulize the protein directly by runing on SDS-PAGE and then by staining with Comassie brilliant blue, if quantity is less then you can stain with silver staining and you can visulize. but if your secreted protein is in very less quantity then you have to concentrate your protein by any of the method, like TCA precipitation or by ammonium sulphate precipitation or any other method and then you cun on SDS-PAGE and can visulize on the gel.
best of luck
regards
aftab
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