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Western blotting: Principles and methods

Cytiva
16 Jan 2023

Since its first description in 1979, Western blotting has become one of the most widely used methods in life science research. Whether you’re a beginner or an expert, this handbook from Cytiva is meant to guide and inspire. Here, Cytiva shares the latest expertise and technology to support your success from sample preparation to analysis. It describes the theoretical and practical aspects of the technique along with useful hints and tips. Over the past decade, improved detection methods and software have brought quantitative analysis to Western blotting. Here, Cytiva gives examples of protocols that can help you get even more quantitative data from your Western blots.

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Flexible imagers with automated features for sensitive, accurate chemiluminescence and fluorescence detection. To support your electronic data management, an optional GxP module is available, designed to meet increasing requirements of regulations like FDA 21 CFR part 11 and EU GMP Annex 11.

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AntibodiesAntibodies are used in techniques such as confocal and fluorescence microscopy, flow cytometry, ELISA, ELISPOT, immunohistochemistry, western blotting and immunopreciptation. Select specific antigen reactivity, high specific affinity, low non-specific binding, monoclonal or polyclonal, primary or secondary antibodies and associated conjugates such as an enzyme or dye for visualization.Western BlottingWestern blotting equipment is used to transfer and identify specific proteins within a sample, reveal protein modifications, as well as give a semi-quantitative estimation of their concentration. Western blotting equipment includes all apparatus necessary to transfer proteins from gel to membrane and subsequent processing steps. Protein transfer can be performed by electroblotting with wet, semi-dry and dry transfer systems onto nitrocellulose and PVDF membranes. Blocking, washing and labeling of membranes follows, involving buffers, blocking reagents, blotting / incubation trays, labeling reagents, immunoblotting assays, antibodies and conjugates. Automated equipment for these steps is available to accelerate your lab workflow. Finally, detection and imaging of proteins can be conducted using gel documentation and imaging systems. Find the best western blotting equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.

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Western blotting: Principles and methods