P53 problems

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TxstateBiochem
TxstateBiochem's picture
P53 problems

 big hinderance for me right now. Im following a number of protocols but nothing seems to be working for me. I have two issues with my protein P53. I can either generate active P53 but when I run an SDS the sample is horribly smeared and contaminated. On the other hand I can generate pure P53 very clean but inactive. Im not sure which would be an easier route I think cleaning the active protein over activating the purifying would be less legnthy but I cant figure out how. Does anyone have any ideas. Right now Ive been seperating it through a column chromatography using different concetraion of a salt laced buffer on a DE52 column but no matter the protein runs off with so many other elements. Im trying to get it pure so I can run alternate tests on teh protein so I can submit an abstract for the upcoming ASBMS.

Thanks in advance.