ResourceLife Sciences
Avoiding Housekeeping Protein Detection Saturation
14 Jul 2013Reliable western blot data require detection of the target and loading control proteins in the linear dynamic range. Many published western blot images show saturated protein bands, especially for the housekeeping proteins (HKP) including ß-actin, ß-tubulin, and GAPDH, indicating that they were not measured in the linear dynamic range. Quantitative analysis based on this type of data is not reliable. This application note describes common ways of avoiding signal saturation for the detection of HKP loading controls.
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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 QuantificationThe detection and quantification of proteins in a sample is vital across life sciences, pharmaceutical research and clinical diagnostics, and a variety of equipment is available to scientists to simplify the workflow. Proteins of interest can be easily labeled and detected on light-based detection instruments. Immunoassay kits allow you to identify a specific protein of interest and protein detection beads or antibody microarrays allow you to identify multiple specific proteins at once. Protein interactions and enzyme activity can also be monitored with protein-protein interaction assays. Additionally, the biophysical characterization of proteins is made easy with biokinetic analyzers. Find the best protein detection and quantification products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.Chemiluminescence