Western blotting, also known as immunoblotting, is a commonly used and acknowledged technique for detecting protein expression levels in cell or tissue extracts. This approach detects protein levels in biological samples by attaching an antibody to a specific protein of interest.
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The precise binding between an antibody and its target protein epitope enables the identification of extremely specific amino acid sequences inside a protein. Antibodies can also identify specific post-translational modifications (PTMs) on proteins.
Phospho-specific antibodies have been utilized to identify components of specific signaling pathways and to study changes in phosphorylation processes in a variety of biological situations.
Researchers have produced antibodies specific to different PTMs, allowing them to monitor changes in a protein’s acetylation, methylation, and ubiquitination status.
Cell Signaling Technology (CST) experts perform over a thousand western blots per day to validate existing and new antibodies.
The western blot protocol, which CST has been optimizing for over two decades, may be found near the end of the eBook and is also accessible online, allowing you to replicate the process and achieve reproducible and dependable results.
This eBook highlights the key steps in the Western blot protocol and demonstrates how adjustments to the protocol can affect the final result. It also discusses the importance of using a well-validated antibody in your Western blot investigations.
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