b'Western blot detectionThe last step in the western blot workflow, after the separation of proteins by PAGE and their transfer from gel to membrane, is detection. In this step, primary antibodies specific to the protein of interest bind the protein on the membrane. With a variety of detection techniques to choose from (chemiluminescent, fluorescent, or chromogenic), you can select a technology to match your experimental requirements and the instruments you have available. Quick visualization or precise quantitation, single-probe detection or multiplexing: we offer a range of reagents and instruments for western blot detection and subsequent analysis. Blocking and wash buffers Verified western blot antibodiesSensitive substrates Imaging and analysisManual vs. automated western blot processingTraditionally, probing a western blot prior to data visualization involves a series of manual steps, many of which are individually short but collectively require significant hands-on time. We offer instruments to automate some of these tasks, tremendously decreasing hands-on time. Automated western blot processing using the Invitrogen Bandmate Automated Western Blot Processor or Invitrogen iBind or iBind Flex Western Device requires only 1530 minutes of setup and no additional hands-on steps before you can move to final detection (Figure 3). Workflow for traditional manual processingPrimarySecondary Prepare buffers and antibody solutions Blockantibody3x washantibody5x wash(2060 min) incubation (5 min each) incubation (5 min each)(124 hr) (1 hr)Workflow for Bandmate processor automated processingPrepare and load blocking buffer, Steps automated with the Bandmate processorwash buffer, and antibody solutionsWorkflow for iBind Western Devices processingPrepare and load Load iBind Card andprimary antibody,Steps automated with the iBind Devices (3 hr)transfer membrane secondary antibody, and wash solutionsFigure 3. Comparison of workflows for traditional manual vs. automated western blot processing. The traditional workflow requires more than 10 hands-on steps and at least 4 hours before final detection can occur. Automated processing using the Bandmate Automated Western Blot Processor or iBind or iBind Flex Western Device requires only 1530 minutes of setup and no additional hands-on steps before final detection. 64 Western blotting thermofisher.com/proteinbiology'