I expressed an extracellular protein and I have obtained good readings on protein assay as it is GFP-tagged. However, I do not obtain any bands for the protein on the SDS PAGE. What could be the possible solution?
Dear Abdullah Ahmad
for any bands in SDS-page do you mean any band at the expected MW? or any bands so empty line?
can please share the SDS-page picture?
best
Manuele
Do you have a positive control for staining, such as another protein, to check for a problem with the staining procedure?
This might arise from when you add your sample or might be because the proteins in your sample were not given enough negative charge. Generally low concentrations of protein could lead to this.
Have you quantified these proteins in your sample? You can add a specific concentration of protein (100ug) to your SDS-PAGE gel to exclude the possibility of low protein concentration in your sample.
Another question, what concentration of separation gel are you using? 12.5%? What size protein in kDa do you want to evaluate?
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