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Quantitative Western Blot analysis is accurate only if the target protein and internal loading control can both be detected in the same linear range. Read this protocol to find out how to determine this combined linear range.
Housekeeping proteins (HKPs) are routinely used for Western blot normalization. But before using an HKP for Western blot normalization, it is critical to validate that its expression is constant across all samples and unaffected by the specific experimental context and conditions. Read this protocol to understand how to do this.
Expression of common housekeeping proteins is known to vary in response to certain experimental conditions. This protocol describes how to use a housekeeping protein for Western blot normalization and quantitative analysis.
Total protein detection is becoming the "gold standard" for normalization of protein loading. REVERT Total Protein Stain is a near-infrared fluorescent membrane stain used for total protein detection and normalization. Learn how to use REVERT Stain for normalization in this protocol.
Replicate measurements are critical for quantitative Western blot analysis. Replicat4e samples confirm the validity of observed changes in protein levels. Learn what biological and technical replicates are and how to perform them effectively.
Traditional, radioactive EMSA protocols can be easily adapted to near-infrared fluorescence EMSA detection by using IRDye® end-labeled oligonucleotides and imaging with the Odyssey CLx or Classic Infrared Imaging System, providing a safe and sensitive alternative.