I would like to know the presence of Insulin mimicking peptide from plant sources, Preliminary I am trying to confirm it by running SDS PAGE. But I am not able to run the Insulin standard. What has to be used as standard. And also preparation of standard. If any one have Idea Pls share.
Hello Raghavendra, Please read the attached documents. i think they will help. Good luck
I have previously experienced that peptides are difficult to be analysed by notorious SDS-PAGE (our unpublished observation at the National Children's Medical Research Center, Taishidou, Setagaya-ku, Tokyo, Japan). Lauroyl or Dodecyl residue (C12 chain) is not sufficient to dissolve hydrophobic molecules. Further, hydrophobic protein such as insulin is difficult to electrophorese (please see file; IEF for hydrophobic proteins, which uses NP-40 (octyl-phenyl chain). Therefore, I have used Brij-58 (palmitoyl chain) in HPLC-SEC analysis.
Therefore, I have developed HPLC methods applicable to hydrophobic proteins.
1) RP-HPLC method (please see file; Insulin RP-HPLC).
2) HPLC-Surfactant-SEC method (please see file; SEC column 300 A silica). This SEC eluent is also applicable to small molecular-mass proteins using 100 A and/or 50 A pore-sized Diol-type silica gels.
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