[ccp4bb] SP Sep HP tight binding of proteins

Dear All, We used SP sepharose high performance as second stage Ion exchange chromatography for polishing the product. We did get pure product but yield obtained was mere 25%. Our protein has a pI of 5.5 - 6.0 and we had used 25 mM Na Acetate buffer pH 4.5 for loading and same buffer with 1M NaCl

Re: [ccp4bb] SP Sep HP tight binding of proteins

Hi Meg, If the bigger peak is appearing when you wash with 1M NaOH, i think your protein is precipitated on the the column. If you have your protein relatively pure after the precedent step, maybe you can try different buffer. You can probably find a better buffer than this one. If your

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