The flour I am testing is quite coarse and the samples are already quite hard to dilute. I tried different dilutions, but after 5 min heating they still become partly solid (like soft ball of dough in the excess water) l suppose that it's because sample is 70% starch and it simply gets gelatinised. However, I did manage to scrape small amount out of "dough" part of the sample and got some results. Are the samples still reliable in this case? Or samples must be liquid to see the real picture?
https://www.researchgate.net/post/Why_did_my_samples_solidify_in_SDS-Gel_Electrophoresis
Hi Dinara,
If starch is really the cause of precipitation maybe you can try treating your sample with some amylase before conditioning it with the sample buffer (take the same quantity of amylase alone as a control). This may also help properly extract the proteins from your flour sample.If proteins are the cause of precipitation you can try to condition the sample you wish to load with 6 to 8M urea (not guanidine HCl since it precipitates with SDS). Adding fresh Beta ME to your sample before loading may be helpful as well. In any cases I would advice not load anything containing precipitated material (starch on any protein material) on your gel since this will affect the quality of your migration.
Good luck.
Hi Dinara,
Use different solvents to extract the protein and compare the results.
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