Contributor:SupryaJayadevDate:September21,1993Resinpreparation:1)Measureoutappropriatevolumeofresinintoa50mlconicaltube.-->NOTE:1mlofDEAE-Sephacelshouldbindapproximately100mgBSA.2)Removeethanolfromresinbywashingresin5Xwithamaximalvolumeofice-coldwatereachtime.3)Washresinonetimewith1MTris,pH7.6.4)Washresin2timeswithlysisbuffercontainingnoDMSOorATP.-->Equilibrateresinwithbufferfor5minuteseachtime.5)CheckthepHoftheresintomakesureitisatornearpH7.5.6)Addsampletoresinandequilibrate(withcontinualmixing)for1to2hours.Columnpreparationandrun:7)Packresinincolumn,drainingdiluentconstantly.-->MAKESUREthatnobubblesforminthecolumnmatrix!!8)Washcolumnwith5columnvolumesoflysisbuffercontainingnoDMSO!!-->NOTE:DMSOstripsproteinsoffofthecoulumn!9)Runa40mlsaltgrADIentcollecting1to2mlfractions:-Startat0mMNaClandendwith500mMNaCl.10)AssayproteinpeaksforSMaseactivity.
