Aggregation of membrane proteins

[Andreas Buhr]

In article <36s16b$k8n at matt.ksu.ksu.edu>, maraman at ksu.ksu.edu (Dayanidhi
Raman) wrote:

> Hi, I am trying to purify a membrane protein using antibody affinity chromatography.  After elution I test them on SDS-PAGE and silver stain them to check for  purity.  But the proteins seems to aggregate & never enters the stacking gel.   If at all it enters into the separating gel it runs a little high than where it is supposed to be.  I do have detergent (nonionic) in the elution buffer.  Does anybody had a similar problem & know how to troubleshoot?.

We had the same experience with IIglc of E. coli. It does not enter the
SDS-PAGE if one boils the samples prior to loading but it does if one omit
boiling.

Good luck
Andreas