Being able to interrogate protein phosphorylation events and aberrant phosphorylation levels is critical to our understanding of various biological and pathological processes. However, detection of phosphorylated proteins in western blots can be challenging. These ten tips will help you generate robust data and cleaner blots.
Normalization of western blot data is very crucial in quantitating proteins. Normalization of band intensity of proteins of interest with the band intensity of housekeeping proteins (HKP) is routinely done in labs. But more and more journals question the validity of using HKPs in normalization and are demanding more validations and tests. An alternative method is to use total protein normalization (TPN) for normalization. Explore the TPN option and see how you can get reliable western blot data easily.
Introducing data normalization into your western blotting workflow may sound unnecessary but it has proven to be a great way to ensure great results. Follow along as we break down some of the main takeaways from the first chapter in Dr. Oh’s popular wester blotting webinar series, including a unique method to ease normalization woes and worries.