Despite the fact that antibodies are one of the most frequently used tools in life science and clinical research, there are no universally accepted guidelines to ensure that they have been properly validated. Learn about the issues facing researchers and what antibody suppliers and researchers can do to ensure that the research being carried out is trustworthy and is helping to improve overall data quality.
Western blotting using fluorescently tagged detection antibodies is a technique that is gaining in popularity. However, adopting a new technique can be challenging. Use the tips provided in this article to get started and see how fluorescent western blotting can enhance your scientific research.
Find out why fluorescent western blotting is gaining in popularity and how it compares to more widely used methods, such as chemiluminescence. From easier multiplexing to brighter fluorophores to easier method development, learn about the many advantages of fluorescent western blotting.
Immunoprecipitation (IP) experiments and the detection of IP samples by western blotting can be quite challenging at times. The tips in this article help you set up and optimize your overall IP experimenta design.
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It is imperative to analyze both the transcriptional and translational profiles for a complete understanding of the functioning of genes. Here we describe a simple protocol that enables parallel analysis of both RNA and protein from single cell culture lysates. See how this workflow could save time and money and yield more reliable data than the traditional protocols used.
Picking the right gel for your application is not easy. It depends on so many factors, such as the purpose of your application, the size of your protein, what kind of buffer you need to have in your sample etc. Here we provide a guide for you to choose the best Bio-Rad gel based on all these factors.
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.
Interference from IgG heavy (~ 50 kD) or light (~25 kD) chains is very common when trying to visualize bands of immunoprecipitated proteins. If your band of interest has a molecular weight close to these, it becomes really hard to distinguish between the IgG and the band of interest. How could this be avoided? Bio-Rad’s new TidyBlot™ Western Blot Detection Reagent accomplishes this by binding exclusively to the native nondenatured antibodies rather than any IgG in the sample. Explore the performance characteristics of this new reagent.
Not all antibodies are created equal. Some have to work hard to qualify to become Bio-Rad’s PrecisionAb™ Antibody. Here is a diagrammatic representation of how the qualification criteria are determined.