I intend performing the partial purification of a recovered peptide using ultra centrifugation and ultra filtration, followed by determining its molecular weight by sds page. The issue is with the sds page system, since despite following the tricine sds page, there is no band observed. How do I trouble shoot the problem?
The problem may be related to both the concentration of protein in the preparation for PAGE and protein loss due to the concentration on the membrane surface during cleaning. Can you clarify the molecular weight of your peptide?
Staining of the proteins is dependent on amino acids composition, so I think you should check amino acids composition of your peptide. If it does not contain positively charged amino acids it will not stain well with Coomassie stain. Then maybe you should try alternative method of staining.
The peptide concentration could be an issue. Perhaps you can try con cryoconcentrate your sample first.
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