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Unformatted text preview: same as above for Sephadex G-150! enzyme in first fraction so PH of 8 > PI or PI <PH of 8 CM-Sephadex is a cation exchanger positively charged proteins will be held on the ion exchanger and -vely charged proteins will wash off. At PHs above the PI. of a protein, the protein will carry a -ve charge and below PI will carry a posoitive one enzyme was in first fraction of column 3 so PI >PH it must be -vely charged so in electrophoresis it goes to the +ve pole ( and depending on it's MW. the distance tot he origin will be determined) urea is powerful protein denaturing agent. it will decrease MW of LDH to somewhere close to 400 so it will take longer to elute urea is uncharged and it will not affect the charge fo the protein .PIBNBE!,BEIJN! 5";!/JDL!8JOUFST...
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- Winter '11