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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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big problem in GST Protein purification
Posted by: Nguyen (IP Hidden, New member, 2)
Date: January 10, 2008 11:48PM
Hi every body!
Recently I was success in joining 2 genes and cloning them as single gene in pGEX-4T1 vector for expression. Although this protein was expressed in soluble form but its amount was quite small. And I already sequenced its and know that its sequencing is Okay ( GST-Gene), besides I check by Western Blot using GST anti as 1st antibody, and the result means that my protein already bind with GST. But problem is when I purify it using Glutathionine 4B sepharose column. My protein can¡¯t bind with resin and nothing in elution fractions. I already check purified ability of this column by run protein from pGEX-4T1 empty vector, and everything is okay. Everybody, have you ever met this condition like me? Please recommend to me how to solve this problem. Thank you very much -NGUYEN-
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