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About GFP- tagged proteins in western blot


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#1 micron

micron

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Posted 29 January 2010 - 02:03 PM

Hi everyone,

I recently bought Adv virus containing GFP fused with my protein of interest. When I did the western blot I saw the shift in the molecular mass of the fused protein. The fused protein should come around 50KDa in western blot but the band appeared art 120kDa. I don't know how this is possible? and will this affect the expression of our protein of interest.

I will appreciate your help in this matter.

Thank you

#2 bob1

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Posted 31 January 2010 - 03:42 PM

Proteins don't necessarily migrate as they are predicted to. Having said that: Are you sure it is your band - detected with both your protein specific antibody and anti-GFP antibody? It is possible that the GFP is blocking the epitope of the protein of interest, so what you are seeing is a non-specific band.




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