AmeriDx® Western Blot Services

The Western blot is an immunological assay to detect target antigen (protein) at the specific position on SDS-PAGE gel, the distance of the position from the gel’s bottom is proportional to it’s molecular weight. With right protein standard, the target protein's molecular weight can be calculated from the western blot image by commercial software or open source software like imageJ.  Similar to the ELISA, it requires an antibody specifically recognizing the target protein, and an enzyme (usually HRP or ALP) conjugated with the antibody to break down the substrate and generate signals which can be viewed by naked eye or fluorescence reader or chemilluminescence reader.

Classic western blot was introduced by Harry Towbin at the Fridrich Miescher Institute in Basel, Switzerland, it has been widely used in protein analysis {Towbin, 1979 #1648}. The basic steps are 1) sample processing; 2) separation of the protein on SDS-PAGE gel; 3) protein transferring to Nitro-cellular membrane or PVDF membrane; 4) blocking of non-specific sites binding to detection antibody; 5) primary antibody incubation; 6) incubation with secondary antibody conjugated with HRP; 7) colorimetric detection of HRP bound to membrane (Fig 1).

Fig 1. Western Blot Schematic Illustration.

America Diagnostics offers reliable and reproducible Western blot services, and free your hand to data analysis and manuscripts writing and let your smart brain focus on key scientific issues. We would be more than happy to offer a trial run for you if antigen and antibody is provided before running the real sample. Please feel free to contact us via email at info@ameridx.com or a quick phone call at (858) 282-8567.