Back in January Judith Airey asked a question about efficiency of IEF gels.
I am having a similar problem and I missed the responses so I hope you
don't mind me asking a similar question. I am a beginner with 2D's and
would appeciate any help.
As with Judith I am finding that the amount of protein present at the end
of the 2nd dimension run is much less than expected. I am using the
Pharmacia IPG strips for the 1st dimension run.
My sample is a mixture of membrane proteins in CHAPS detergent. The sample
has protease inhibitors present so proteolysis shouldn't play a major part.
Could the solubility of the membrane proteins be the problem, perhaps with
the protein precipitating out during the 1st dimension run? If this is a
potential problem has anyone got any useful suggestions for increasing the
solubility of the sample, or any ways to improve efficiency generally.
Any comments would be gratefully received
Thank you very much
Amanda HAll
Amanda Hall
Professional Officer
Newcastle Protein
University of Newcastle
ph 02 4921 7299
email: mbah@medicine.newcastle.edu.au