hello All,
I am trying to produce Human TGFB1 in tobacco by using Agrobacterium mediated transient expression. The expression is very good, protein is perfectly soluble in buffer. In the literature I have found that free form of TGFB1 is highly hydrophobic and easily interact with the E-tubes or other hydrophobic surfaces and that's why when i remove all the tag from rest of the protein, TGFB1 yield is very low or sometimes all is lost. 2nd point is, commercial TGFb1 is reconstituted in vey low pH buffer (~3-4, in 10mM HCl or 10 mM Citric Acid) while i am using tag removal buffer the pH of which is around its pI value 7.5.
tag removal enzyme is not efficient at that low pH (3-4). In order to recover protein any suggestion will be highly appreciated.
thanks in advance, please do share your experience
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