As a laser-based NIR fluorescence scanner platform, the Sapphire™ NIR-Q enables total protein normalisation for more efficient and better quality quantitative Western blotting workflows. Combining all the advantages of the highly sensitive, low background and fast-scanning Sapphire™ NIR, the Sapphire™ NIR-Q has two channels for multiplex protein detection and a third channel for total protein detection.
Benefits of using the Sapphire™ NIR-Q biomolecular imager
• 658 laser and 784 laser (both suitable for detection of modified or unmodified proteins) and a 520 laser for total protein normalisation and dye flexibility
• Detect two proteins and normalise to a HKP loading control
• Detect two proteins and normalise to a total protein stain
• Detect two proteins and normalise to a total cell stain
• Sapphire Capture software for high quality yet simple image capture
• High resolution image capture - down to 10 microns
• High sensitivity for lower limits of detection (femtograms)
• Ultra-wide dynamic range for accurate quantification
• Low and high abundance samples quantified simultaneously
Total protein normalisation
General housekeeping proteins have been used widely for normalisation when quantifying Western blot (WB) bands. However, journals are now moving towards a preference for total protein normalisation for quantitative WB. Adding total protein staining to the Western blotting process can complicate an already time-consuming, multistep workflow. It may also require stripping and reprobing of the blot if the scanner does not contain enough channels to image everything at once.
For researchers needing to scan only in the NIR fluorescence range, the Q-module addition to the Sapphire™ NIR biomolecular imager adds a third channel for visualising total protein staining (with Cy3 dyes or AzureRed fluorescent protein stain) and then probing for two proteins using the two NIR channels.
Total protein normalisation with AzureRed fits easily into any Western Blotting workflow
Request a technical presentation or find out more by contacting our Sapphire specialists here.
Material available for download
Application note: Accurate Western Blot Normalisation with AzureRed Fluorescent Protein Stain
Note: product availability depends on country - see product detail page.