ResourceLife Sciences
Phosphorylated Protein Detection is More Efficient by Fluorescent Western Blot
21 Nov 2017This application note demonstrates the convenience and accuracy of fluorescent Western blotting for the detection of phosphoproteins in comparison to traditional chemiluminescent assays. A protocol for the visualization of modified and unmodifed proteins simultaneously is outlined, which improves the speed and accuracy for multiplex detection of proteins that requires no blot stripping.
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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.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.BiomarkersBiomarkers are biological markers which can be measured and evaluated to indicate a biological state. The use of biomarkers in research and diagnosis can indicate a normal or disease state or drug response of cells / tissues. Biomarkers include genetic markers, cell surface markers such as antigens, antibodies or receptors and secreted molecules such as cytokines. An assay system is required for identification of biomarkers. :ImmunofluorescenceFluorescence Based AssayFluorescence based assays are widely used in life science research and high-throughput screening to measure a broad range of cellular activities.Protein DetectionProtein detection techniques are essential for understanding protein expression, localization, and function. Methods such as western blotting, ELISA, and mass spectrometry are commonly used in research and diagnostics. Browse our peer-reviewed product directory to find the best tools for protein detection, compare products, check reviews, and get pricing directly from manufacturers.Cancer ResearchCancer research aims to understand the mechanisms of cancer development and progression to improve prevention, diagnosis, and treatment. From molecular biology to clinical trials, research spans a wide range of disciplines, including immunotherapy, targeted therapies, and drug discovery. Explore the best cancer research products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.MultiplexingMultiplexing refers to the ability to measure multiple targets or analytes simultaneously in a single experiment. This technique is valuable for high-throughput screening, diagnostics, and complex assays, as it increases efficiency and data quality. Browse our peer-reviewed product directory to find the best multiplexing products, compare tools, check reviews, and get pricing directly from manufacturers.