|Petite Kit Includes||Quantity|
|LumiGLO® Reagent A (20X)||1 x 5 ml|
|Peroxide Reagent B (20X)||1 x 5 ml|
|Small Kit Includes||Quantity|
|LumiGLO® Reagent A (20X)||1 x 25 ml|
|Peroxide Reagent B (20X)||1 x 25 ml|
(a) Wash membrane-bound HRP (antibody conjugate) three times, for 5 minutes in TBS/T.
(b) 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).
(c) Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film.
Prepare solutions with Milli-Q® or equivalently purified water.Wash Buffer (TBS/T): 20 mM Tris-HCl (pH 7.6),137 mM NaCl and 0.1% Tween-20Advantages of the Phototope® Western Detection System-Sensitivity: Detection of sub-picogram amounts of protein is routine with good primary antisera.-Speed: Less than 1 hour is required for the entire detection procedure. Exposure times are seconds to minutes.-Multiple Exposures: Light is emitted at a constant rate for several minutes, so you can perform multiple exposures to optimize signal intensity. Re-exposure at a future date is achieved by simply adding more reagent.
Store at 4°C.
There are six basic steps in the Western blotting procedures with the Phototope®-HRP Western Blot Detection System.
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