ResourceLife Sciences
Application of the Bio-Plex Cell Lysis Kit in Western Blot Analysis of Phosphoproteins
7 Mar 2017The Alliance for Cellular Signaling (AfCS) is charting interactions among cellular signaling molecules to produce theoretical models that will describe how cells respond to stimuli. The objective of this application note is to reduce the labor involved and substantially expand capabilities by transitioning to Bio-Rad’s Bio-Plex phosphoprotein assays, which employ Luminex xMAP technology and Cell Signaling Technology’s phosphorylation site-specific antibodies.
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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.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.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.Monoclonal AntibodiesMonoclonal antibodies (mAbs) are lab-made molecules that can bind to specific antigens on cells, making them highly effective in treating cancers, autoimmune diseases, and infections. These targeted therapies are transforming the treatment landscape for many conditions. Browse our peer-reviewed product directory to find monoclonal antibodies for research and clinical applications; compare products, check reviews, and get pricing directly from manufacturers.Cell BiologyCell biology studies the structure, function, and behavior of cells. Understanding cellular processes is essential for research in areas such as cancer biology, stem cell research, and neurobiology. Techniques like flow cytometry, microscopy, and cell culture enable researchers to explore cellular mechanisms in detail. Browse our peer-reviewed product directory to find the best cell biology tools and equipment, compare products, check customer reviews, and get pricing directly from manufacturers.Cell SignalingPhosphorylationProtein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.