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I am trying to express a FAD containing enzyme of mycobacteria in E.coli. I am able to purify the protein which is slightly yellow in color but it seems that my FAD is all unbound to the protein....
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I want to prepare reduced form of NADPH. I am using 100mM of Tris (pH8.0) to make the buffer but the absorbance is showing to be 1.6 at 340nm which should supposed to be 0.6. can someone please...
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I tried to do same with membrane fraction. It expressed in membrane but somehow not working in periplasm
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