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in this video we will show you how to prepare every block blocking buffer to block your membrane and perform antibody incubations in your western blotting experiment every block blocking buffer is an optimized buffer for blocking membranes prior to immunodetection this buffer enables effective blocking in only five minutes without compromising sensitivity and is compatible with all detection methods to block the blot add enough every block blocking buffer to cover the entire membrane incubate for five minutes at room temperature with gentle agitation dilute your primary antibody directly into fresh full strength every block buffer following the antibody manufactureramp;#39;s directions for dilution strength pour off the buffer from the previous blocking step then add the primary antibody buffer mix incubate the membrane for one hour with gentle agitation at room temperature wash the membrane five times with tbst for five minutes each with agitation dilute the secondary antibod