Labeling CGC 649 Peptide (P6014)
Protocol
- Dissolve 10 nmoles of peptide in 20 µl water to a final concentration of 500 µM.
- Thaw the protein containing the C-terminal thioester.
- Mix the following components:
Component Volume Final Conc. Peptide Solution (500 µM) 2.5 µl 100 µM IPL Buffer (10X) 2.5 µl 1X Protein (1 mg/ml) 10 µl 0.4 mg/ml dH2O 10 µl Total Volume 25 µl - Incubate at 4°C overnight.
- Add 12.5 µl of 3X SDS-PAGE Sample Buffer.
- Heat at 95°C for 5 minutes.
- Load approximately 150 ng protein/lane.
- Transfer protein to a membrane (e.g., nitrocellulose).
- Visualize using a fluorescence scanner at the appropriate wavelength.