Home Protocols Labeling CGC 649 Peptide (P6014)

Labeling CGC 649 Peptide (P6014)

Protocol

  1. Dissolve 10 nmoles of peptide in 20 µl water to a final concentration of 500 µM.
  2. Thaw the protein containing the C-terminal thioester.
  3. 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  
  4. Incubate at 4°C overnight.
  5. Add 12.5 µl of 3X SDS-PAGE Sample Buffer.
  6. Heat at 95°C for 5 minutes.
  7. Load approximately 150 ng protein/lane.
  8. Transfer protein to a membrane (e.g., nitrocellulose).
  9. Visualize using a fluorescence scanner at the appropriate wavelength.