Our featured application notes below provide further details. The Design of a Quantitative Western Blot Experiment Read a summary about a complete western blot workflow with a focus on sample preparation and data analysis for quantitative western blotting. Utilizing this substrate-free assay, researchers can achieve equivalent sensitivity to traditional chemiluminescence assays while consolidating western blot, ELISA, and other applications onto a single reader. Bottom panel: Quantification of AhR protein levels from Western blot analysis (n 3). Each type of detection has advantages and disadvantages that need to be considered when selecting a method.Īnother method that we have introduced the ScanLater™ Western Blot Detection System, enables first-of-its-kind western blot detection in a multi-mode microplate reader platform. For example chemiluminescence can be detected using x-ray film or digital imaging equipment while a fluorescent secondary antibody requires a fluorescence imager. These methods are described below and require different equipment for detection. These problems stem from a low-dynamic range of detection and the difficulty in accurately determining the limit of detection 6, 7. There are several western blot methods to consider depending upon the secondary antibody used, detection of the target protein may be colorimetric, chemiluminescent, or fluorescent. Chemiluminescent western blot data, derived from film-based detection, poses distinct challenges in producing quantifiable, reproducible data. Related articles Enrichment and Detection of Wnt Proteins from Cell Culture Media SDS-PAGE and Western Blotting: Basic Principles and Protocol Size Analysis. HIGH-THROUGHPUT, HIGH CONTENT SCREENING.PROTEIN DETECTION, QUANTITATION, ANALYSIS.NUCLEIC ACID (DNA/RNA) DETECTION & ANALYSIS.
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