There can be two suggestions from my side:
1. Prepare the sample freshly, heat properly for denaturation, and do not store it for longer.
2. Check the proper concentrations of Sample buffer most importantly the denaturing agents B-Mercaptoethanol or DTT (one agent can be used at once and not both).
Maybe is the protein concentration.
Did you cuantify it before your well load?
When we was in our western blot standarization phase, we obtained SDS-PAGE with smears, and we determine that 50 ug - 80 ug of protein concentration is a great range for our electroforesis.
With 50 ug we get SDS-PAGE like the photo below.
Other consideration is your SDS-PAGE quality, maybe didn´t have a good polimerization.
Regards
if you really need that much protein/lane, try Laemmli sample buffer with with 6 M urea. But also check your actual SDS and beta mercap concentrations.
Also, if you have any nonionic detergent (triton etc) in the previous step make sure you have a good molar excess of SDS (ie use 5x etc).
There can be two suggestions from my side:
1. Prepare the fresh homoginate buffer (all essential buffer).
2. Fresh Tris buffer use (pH maintain required your protocal).
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