Western blotting has long been a core technique in the biochemistry of protein analysis. In fact, it is often a confirmatory method for many complimentary protein analysis methods like ELISA and mass spectroscopy, particularly when used in clinical settings to aid in diagnosis.

However, the western blotting service blotting procedure is well known to be labor intensive, expensive and requires a minimum of 10 mL of sample for one assay. The process is lengthy, with 8-20 h required for gel preparation, separation, transfer, and multiple incubations. This is time consuming and may decrease the reproducibility of the results. Additionally, the western blot is not readily miniaturized, which limits sensitivity and increases materials consumption.

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The detection step of a western blot is dependent on the label used to detect antigens. The most common method of detection is indirect using an enzyme- or fluorophore-conjugated secondary antibody to detect the target protein on the membrane. Alternatively, direct detection can also be performed by first using an unlabeled primary antibody to bind to the target antigen and then directly detect the resulting signal with either chemiluminescence or fluorescence.

Quantitative western blot data is increasingly being utilized in research to more accurately delineate the biochemical attributes of proteins, and upstream cell-signaling processes important in both normal and disease physiology. This includes accurate protein abundance determinations, 3-dimensional modeling of protein complexes, and phosphorylation stoichiometry of key proteins in signaling pathways.