Western blotting is an important technique used to identify specific proteins from a complex mixture of proteins extracted from cells. The technique uses three elements to accomplish this task: (1) separation by size, (2) transfer to a solid support, and (3) marking target protein using a proper primary and secondary antibody to visualize and quantify. [...]
The use of primary antibodies is essential for Western Blotting procedures. These antibodies are sold in small volumes (larger volumes are also available) and can be expensive. The typical starting dilution of primary antibody is 1:1000 unless there is a recommended starting dilution. Initial testing to optimize the antibody concentration could be a series of [...]
Western Blotting is used for the detection of a specific protein using immunodetection. The process involves the immobilization of a protein of interest to a membrane, PDVF or Nitrocellulose. Once the protein is bound to the membrane it is then probed with a primary antibody and a secondary antibody conjugated to an enzyme (HRP or [...]
Bad Western blots. It happens! Use our Western blot troubleshooting guide to help you sort it out so that your next blot is your best blot.
When the World Anti Doping Agency (WADA) needed an automated Western blot device to test samples from over 10,500 of athletes, they chose the BlotCycler™ for fast, reliable, reproducible results with SAR-PAGE.
When the WADA accredited laboratory in Lausanne, Switzerland needed to analyze over 1000 samples for the 2014 FIFA World Cup, they used the BlotCycler™ for speed of processing and reliable results for their Western blots!
When researchers investigated the optimization of SAR-PAGE, they used the BlotCycler™ for incubation and washing to increase the reproducibility of results and economize labor time and increase signal-to-noise ratio.
When researchers investigating Rapamycin treatment of healthy pigs and cardiac functions, they used the BlotCycler™ Automated Western Blot Processor for increased reliability and validity.
When researchers evaluated the potential use of clone 9G8A for doping control purposes, they used the BlotCycler™ automated Western blot processor to enhance reliability and validity of their findings.
Don't see the protein expression by Western blotting even after trying multiple antibodies and other protocol optimizations? The reason could be as simple as the protein loss from the membrane. Use our PVDF western blot troubleshooting strategy for greater success.
