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EXPRESSION PROBLEM IN GST TAGGED PROTEIN


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#1 anuj

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Posted 19 May 2011 - 11:06 PM

hi, i have a GST tagged clone in pGEX 4T vector. my protein size is 128kda and GST is 26kDa (approx). so finally total size is 154kDa approx. i am trying to express in BL21. i have tried at 24 and 28oC temp with 0.5mM, 1.0mM,3mM and 5mM IPTG conc. i dont find any difference in induced and uninduced samples. but when i pull down with GST beads i find a very high level of GST expression but my protein is not at all expressed. please suggest ..

#2 Andrey

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Posted 20 May 2011 - 01:47 AM

Are you sure that you're expressing from the vector with an insert and not from an empty one?




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