What can protein extraction buffer change in a Western blot?
Chosen extraction buffer can actually change quite a lot, as in this example:
Anti-VDE antibodies used on extracts from:
Brachypodium distachyon, Arabidopsis thaliana, Oryza sativa
Adjustment of a buffer to: 7M Urea, 2M Thiourea, 4% CHAPS, 35 mM Tris pH 8 resulted in deteciton of VDE in two more species: Brachypodium distachyon and Oryza sativa.
If the antibody you are working with, is not giving you a signal on other model species, but the antigen used to elicit this antibody is perfectly conserved in a sequence of a protein from other species, specific adjustments of Western blot protocol are necessary.
Usually lack of detection in another model species may be connected with epitope inaccessibility. Therefore, application of more denaturing conditions can be helpful to resolve such issue.
Left panel no urea in extraction buffer
Brachypodium distachyon, Arabidopsis thaliana, Oryza sativa
Right panel urea in extraction buffer
Oryza sativa, Brachypodium distachyon, Arabidopsis thaliana
Presented result is a courtesy of Dr. Nelson Saibo,Plant Gene Regulation Lab (GPlantS Unit), ITQB NOVA, Portugal