CST, 7003P, 20X LumiGLO ® Reagent and 20X Peroxide

Product Usage Information
Prepare solutions with Milli-Q ® or equivalently purified water. 1. Wash membrane-bound HRP (antibody conjugate) three times, for 5 minutes in TBS/T (20 mM Tris-HCl (pH 7.6),137 mM NaCl and 0.1% Tween-20) 2. Prepare substrate by diluting 20X LumiGLO ® and 20X Peroxide to 1X in water (e.g. for 10 ml, add 0.5 ml LumiGLO ® and 0.5 ml peroxide to 9.0 ml water). 3. Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film.
Storage
Store at 4ºC. This product is stable for 36 months. DO NOT store Reagents A and B pre-mixed. Reagents A and B should be combined just prior to exposing membranes.
Background
Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.
Alternate Names
20X LumiGLO® Reagent and 20X Peroxide; chemiluminescence; ECL; Luminol