Our highly sensitive ECL Western blotting detection reagents are shipped in 2 x 100mL of A and B solutions. Solutions A and B are to be mixed in a 1:1 ratio prior to application. The mixture is then ready for detection of any immobilized specific antigens in chemiluminescent Western blots through horseradish peroxidase (HRP) conjugated antibodies. The kit is designed for a high quality and long lasting chemiluminescent reaction.
Protocol
Wash membrane thoroughly after antibody incubation and before initiating ECL.
Immediately before use, mix solutions A and B in a 1:1 ratio in a low light intensity or dark room. 1-2ml of solution is recommended for every 10cm3 of membrane.
Add the blot to the reagents and incubate for 1 minute while shaking.
Pour out the solution, dab off any excess solution and wrap the blot in clear plastic.
Place the blot into a film cassette and expose to X-ray film in a dark-room setting immediately.