His fused protein - (Aug/27/2007 )
Hi..
I have a protein with 8 His tag, and I have problem with purifying it using Nickl affinity chromatography. at the same time in my westren blot I use anti his antibody (tetra) and I have more than one band. I am thankful for your suggestions.
-hemin-
you have several bands... Well maybe 6his antibody is not that specific. Colocalizes with your protein (with anti-protein antibody). You'll discriminate from degradation of our protein.
8his tag, i've heared regulary of 6his. Why 8 ?
-fred_33-
QUOTE (hemin @ Aug 27 2007, 01:56 AM)
Hi..
I have a protein with 8 His tag, and I have problem with purifying it using Nickl affinity chromatography. at the same time in my westren blot I use anti his antibody (tetra) and I have more than one band. I am thankful for your suggestions.
I have a protein with 8 His tag, and I have problem with purifying it using Nickl affinity chromatography. at the same time in my westren blot I use anti his antibody (tetra) and I have more than one band. I am thankful for your suggestions.
May be you are getting dimers and trimers. Check the molecular weight of the protein on the blot and calculate
-Anjou-