Description

Product Description: ECL Western Blot Kit is a ready to use for chemiluminesecent detection on protein conjugated with HRP on the membrane directly.
Content Table:
ECL Reagent-1 250 ml 6 ml/sheet for 100 assays,8 ml/sheet for 75 assays. At 4?C for 6 months -1 year
ECL Reagent-2 250 ml 6 ml/sheet for 100 assays,8 ml/sheet for 75 assays. At 4?C for 6 months -1 year
Quick Protocol:
1. * Mix an equal volume mixture of ECL Reagent 1 and 2 to give enough solution to cover the membrane (0.15-0.2 ml/cm2). Let the mixture equilibrate for 5-10 seconds. It is ready ECL mixture.
*doing the process under the nature light. Or in dark Room under the red safely light.
2. Place the Western Membrane, washed well, in the ready ECL mixture, and make sure the protein side is up, and covered inside the ECL mixture, shaking at RT for 30-60 seconds.
3. Take out the membrane and to drain off excess ECL mixture with paper towel. Wrap the membrane in saran wrap. Gently remove air pockets.
Above 1-3 is the processing in Dark room or in Room without light.
4. Place the membrane, protein side up, in the cassette; place a film on the membrane to be exposed for 10-30 seconds under the dark or with the red safety light. Then to be developing in the machine.
5. If the signal was too low, you could keep the exposure at RT for 30 minutes. Then to be developed again.
6. If the signal intensity was too high: 1). you could keep the film exposed for 5-25 minute in dark, then to be developed again. 2). you also to dilute and use 1/3-1/2 ECL mixture in wash buffer for your ready western membrane to get the desired results.

References:
1. Tsukagoshi K, et al.: Enhancing effect of phenylboronic acid compounds and their interactions with the diol groups of saccharides in a capillary electrophoresis-chemiluminescence detection system. Anal Sci., 2007 Feb; 23(2): 227-30.
2. Alpeeva IS, rt al.; Luminol-hydrogen peroxide chemiluminescence produced by sweet potato peroxidase. Luminescence, 2007 Mar-Apr; 22(2): 96-6.
3. Semenkova GN, et al.; Chemiluminescence in the peroxidase oxidation of luminol with hydrogen peroxide in various media. Lab Del; 1991(11): 13-5

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ECL Western Blot Detection Kit

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