SuperBlot^TM ECL Western Blotting Substrate Kit (Standard)

Protocol for Chemiluminescent blot development with ECL

Digital Imaging

1. Remove blot from the final wash solution and place on an imaging tray
2. Mix equal quantities of part A and part B solutions being careful not to contaminate solutions by changing pipette tips
   1. For a 10x7.5cm gel we recommend 750µl of each solution
3. Add combined solutions to the blot making sure to cover the entire area.
4. Cover blot with a clear transparent sheet of plastic to prevent evaporation then immediately image.
   1. Be careful to not introduce any bubbles as these will show up in the final image.

Film Imaging

1. Mix equal quantities of part A and part B solutions being careful not to contaminate solutions by changing pipette tips.
   1. For a 10x7.5cm gel we recommend 750µl of each solution
2. Add combined solutions to a sheet of cling film large enough to fit the blot.
3. Remove the blot from the final wash buffer, dab off any excess then place into the ECL solution. Use tweezers move the blot in order to soak it in ECL making sure to cover each side thoroughly.
4. Dab off any excess ECL with filter paper then place into a pocket of clear plastic within a X-ray imaging cassette.
   1. Be careful to not introduce any bubbles as these will show up in the final image.
5. Move to a dark room.
6. Cut a piece of X-ray film to size then tape to the opposing door of the cassette. Close the cassette in one clean motion so that the film and blot are now in contact.
7. Expose the film for an appropriate amount of time.
   1. This will vary depending on the target protein and which primary and secondary antibodies are used.
8. Open the cassette and develop the film.

+4°C (protect from light)

Contains 0.05% ProClin-300