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Western Blotting Protocol

In this video we take you through all the steps in our Western Blot (WB) Protocol so you can replicate our procedure and get reproducible and reliable results. 👉 UPDATED CST Global Contact Page: https://www.cellsignal.com/about/cont... 👉 CST Protocols and Troubleshooting: https://www.cellsignal.com/protocols 👉 Western Blot Troubleshooting Video:    • Western Blot Troubleshooting Guide   👉 Western Blot Tech Tips playlist:    • Western Blotting (WB) Protocols and Tips   👉 Subscribe: https://youtube.com/user/cellsignaldo... Contents: 0:00 Introduction 1:06 Solutions and Reagents 2:11 Sample Preparation and Protein Blotting 4:33 Membrane Blocking and Antibody Incubations 6:07 Protein Detection 6:45 Importance of Following the Recommended Protocol 8:37 Conclusion Western blotting is a widely used immunoassay used by research scientists to monitor expression of a specific protein in a complex cell or tissue extract. It utilizes antibodies that recognize a specific protein of interest or a post-translational modification like phosphorylation, acetylation, methylation, and ubiquitination. At Cell Signaling Technology (CST), we perform thousands of Western blots daily using a protocol that has been optimized for over a decade to develop and validate antibodies with exceptional specificity, sensitivity, and reproducibility. We first list all the the solutions and reagents you’ll need. Then, we give detailed descriptions on how to prepare the sample, perform the protein blot and immunoassay, and detect proteins chemiluminescently. Finally, we describe critical experimental steps in Western blotting and explain how small changes to the protocol, like changing incubation times or dilution buffers, can affect the final outcome of your blot. To learn more about how to generate publication-ready Western blot data, check out our troubleshooting video (   • Western Blot Troubleshooting Guide  ). If you’re interested in alternate protocols, including preparing your sample with immunoprecipitation, re-probing your membrane, and detecting your protein using fluorescence detection, visit Protocols at the Cell Signaling Technology website (https://www.cellsignal.com/protocols). About CST: Cell Signaling Technology (CST) is a different kind of life sciences company—one founded, owned, and run by active research scientists, with the highest standards of product and service quality, technological innovation, and scientific rigor. We are a company of caring people driven by a devotion to facilitating good science—a company committed to doing the right thing for our Customers, our communities, and our planet. https://https://cellsignal.com/about-us All trademarks are the property of their respective owners. https://cellsignal.com/trademarks