Routinely used to study cancer, HIV, sports doping, Lyme disease, mad cow disease, etc., etc., western blots are notoriously unreliable. Variations in technique confound reproducible results. Retraction Watch, a blog that reports on retraction of scientific papers, questions whether western blots are trustworthy at all. What’s going on? Koller et al. writes that variable results [...]
Each antibody-antigen interaction has unique characteristics. Interaction of antisera with protein epitopes in western blotting depends upon a number of factors, all contributing to the final signal/noise ratio. Knowledge and control of those factors allows for modifications and optimization of the procedure. Prepare the membrane For size-related identification of an interaction between protein and antibody [...]
A successful western blot is determined by more than the primary antibodies. Although many researchers believe that a successful western blot is determined by the nature of the primary antibodies, every variable within the assay can influence data quality. For reliable and reproducible quantification of western blot, automation is key. Manual processing can [...]
We recommend you wear gloves and use forceps for reproducible western blot assays Follow these tips for improving the reproducibility of your western blot assay, and for achieving experimental results that are actually publishable. Each antibody-antigen interaction has unique characteristics. The interaction of antisera with protein epitopes in western blotting depend upon a [...]
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.