Hello Jerry, I would be glad to provide protocol suggestions, but it'd help to know a bit more regarding what you are working on. If you're planning to purify material from its native source (i.e. someone's chopped up sittin' muscle?) - this would be tremendously different from purifying protein from transfected CHO, HEK, etc. cells where you can expect reasonable abundance.
Details are very important :) Artem > > Hi, All: > > Although it is off-topic, definitely I think I can get some help > here because we crystallographers are dealing with protein > purification almost every day. > > My protein was expressed in E.coli as a soluble octamer with or > without his-tag revealed by gel-filtration. For the his-tagged > protein, the final product is an octamer after Ni-column and > gel-fil. However, purification of the non-tagged protein results in > a tetramer because of a partially denatured condition. > > When I tested the enzymatic activity , it turns out the tetramer was > active although both of them can be crystallized. Now I am > wonderring whether its native form in human is an octamer or > tetramer. > > I am planning to purify a little protein from human cells to verify > its native form. > > Can anybody recommend some protocols? > > Thanks a million, > > Jerry McCully > > > _________________________________________________________________ > Rediscover Hotmail®: Now available on your iPhone or BlackBerry > http://windowslive.com/RediscoverHotmail?ocid=TXT_TAGLM_WL_HM_Rediscover_Mobile1_042009
