i'm working on protein evaluation in flour of wheat and others legumes.....I'm using SDS-PAGE. However those days (about two weeks ago) i foudd nothing in the gels (no bands), i dont know what's the problem, i regulated the pH of the running buffer, also the pH of others solutions but hopeless.....PLEASE i need idea to resolve this problem :(
Hi Neila Abdi did you notice the amount of sample you added on the gel? May be it might not be enough to show any band and how about sample preparation, was it based on the heating method with sample buffer? (as described from Sambrook et al Molecular cloning manual)
Did you see your marker bands in the gel? Are you sure about your protein extraction procedure? You can load simultaneously some positive control samples which surelyhave has been extracted correctly and able to be observed in gel.
By the way, some suggestions:
Ensure that the protein concentration of each sample is sufficient and accurate.
Not enough SDS in the sample: Without adequate SDS, the proteins are not negatively charged and will not travel through the cell. Ensure adequate SDS concentration.
For more information, please check:
https://www.sigmaaldrich.com/technical-documents/articles/biology/western-blotting/troubleshooting/sample-preparation-gel-electrophoresis.html#No%20Bands%20or%20Gel%20Front
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