I want to quantify my protein extracts that have been solubilized in IEF sample buffer. what do i do? can some one help.
IEF sample buffer contains DTT , ampholytes, CHAPS which interfere with quantification using bradford's method. Reduce the concentration of these agents by diltuing to the compatible concentrations for the method of quantification you are goinf to use.
Try solubilizing protein first in a compatible buffer, quantify and then solubilize in the IEF buffer from next time onwards.
I recommend BCA assay. Below is a link to the reagent (and procedure details).http://www.piercenet.com/products/browse.cfm?fldID=02020101
Thanks let me try it.
Thanks, it is wise to solubilize first in compatible buffer first then solubilze in the IEF buffer.