b'Manual vs. automated western blot processingTraditionally, probing a western blot prior to data visualization involved a series of manual steps, many of which were individually short but collectively required significant hands-on time. Today, instruments are available to automate some of these tasks, tremendously decreasing hands-on time.Manual and automated procedures share three essential steps: blocking the membrane, probing with primary and secondary antibodies, and washing the membrane (Figure 14).Traditional manual processing workflowPrimarySecondary Prepare buffers and antibody solutions Blockantibody3x washantibody5x wash(2060 min) incubation (5 min each) incubation (5 min each)(124 hr) (1 hr)Bandmate processor automated processing workflowPrepare and load blocking buffer, Steps automated with the Bandmate processorwash buffer, and antibody solutionsiBind Western Devices processing workflowLoad iBind CardPrepare and load and transferredprimary antibody,Steps automated with the iBind Devices (3 hr)membrane secondary antibody, and wash solutionsFigure 14. Comparison of traditional manual vs. automated western blot processing workflows. The traditional manual western blot processing workflow requires more than 10 hands-on steps and at least 4 hours before final detection can occur. Automated western blot processing using the Invitrogen Bandmate Automated Western Blot Processor or Invitrogen iBind or iBind Flex Western Device requires only a 1530 minute setup time and no additional hands-on steps before you can move to final detection.29'