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Efficient total protein normalization for quantitative Western blots

While many researchers use general housekeeping proteins to normalize for load when quantifying bands on a Western blot, the past few years has seen a movement towards using total protein staining instead, for more accurate quantitation (see CP Moritz. Tubulin or Not Tubulin: Heading Toward Total Protein Staining as Loading Control in Western Blots. Proteomics. 2017 Oct;17(20). PMID: 28941183.).

However, adding total protein staining to the Western blotting process can complicate an already time-consuming, multi-step workflow. In addition, depending on the type of stain you use and the number of different proteins being quantified, there might not be enough channels in your scanner to image everything at once, necessitating even more steps to strip and re-probe the blot. For example, the LI-COR® Odyssey® CLx has only two channels, which means only one protein can be measured at a time (one channel to image the total protein stain and one channel for the protein-of-interest).

Fortunately, there’s the Sapphire™ NIR-Q Biomolecular Imager. For researchers needing to scan only in the NIR fluorescence range, the addition of the Q-Module to the base Sapphire™ NIR Biomolecular Imager adds a third channel for visualizing total protein staining using Cy3 dyes or our own AzureRed Fluorescent Protein Stain and then probing for two proteins using the two NIR channels. Increase your efficiency while increasing the quality of your quantitative Western blots with the Sapphire NIR-Q.

If you’d like to be even more efficient, the Sapphire™ RGBNIR enables four-channel fluorescent imaging so you can probe for three different proteins and still detect total protein stain without stripping and re-probing.

Experience the speed and simplicity of the Sapphire™ NIR-Q and set up a demo by filling out the form on the right.

 

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Supported Applications

2D-DGE • FLUORESCENT PROTEIN NATIVE GEL • REPORTER GENE ASSAY • QUANTITATIVE WESTERN BLOT • NUCLEIC ACID GEL DOCUMENTATION • ELISA • IN-GEL WESTERN • PROTEIN GEL DOCUMENTATION • PROTEIN ARRAY • IN-CELL OR ON-CELL ELISA • DENSITOMETRY • CHEMILUMINESCENT ARRAY • QUANTITATIVE WESTERN BLOT • ELECTROPHORETIC MOBILITY ASSAY • CHEMILUMINESCENT WESTERN BLOT • IN-CELL WESTERNS • PHOSPHOR IMAGING • TISSUE SECTION IMAGING

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See how efficient total protein normalization is on a Sapphire NIR-Q—set up demo or learn more by filling out the form below.