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Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature. Wash three times for 5 min each with 15 ml of TBST. Proceed with detection Section D. Detection of Proteins Directions for Use: Incubate substrate with membrane for 1 minute, remove excess solution membrane remains wetwrap in plastic and expose to Site rencontre bucarest film. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment.

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