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Jacobs:Protocol NuPAGE Electrophoresis and Western Blotting for Proteins
Revision as of 06:34, 21 July 2008
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06:34, 21 July 2008
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#Incubate membrane for 1hr at room temperation with agitation in anti-goat-HRP conjugate (1:2000 dilution in TBS)
#Wash membrane 3x (5-10min each) with TBS
Mix 1 mL of Detection Reagent 1 and 2 (1 mL of each),
#Pour over transfer membrane and leave on for one minute.
#Dry off with Kimwipe
#Bring to the dark room and make sure the door is tightly closed (no light seeping through), before beginning
#Take out Kodak scientific imaging film and cut in half. Put the other half back in the box.
#Put Kodak film over transfer membrane in cassette and close cassette. Leave for one minute. (Note: Do not expose film to light. Keep lights off until film is in imaging machine)
#Put film into imaging machine to process
#If nothing shows, put the film back into the cassette and leave it in for another 30 minutes. Check back to see if results show.
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