protocols and tips

Analyzing antibody binding to histidine-tagged proteins is not always easy because of inherent problems such as nonspecific binding and loss of analyte response associated with these tagged proteins. Optimization of these factors is key to precisely measuring the binding kinetics of these proteins.

Today many researchers are considering changing their western blot detection method from chemiluminescence to multiplex fluorescence. There are several drivers behind this trend. Most significantly, fluorescent detection allows users to multiplex their western blots, enabling simultaneous detection of several target proteins at once, reducing or eliminating the need to strip and re-probe. Other benefits of fluorescence include better dynamic range, more quantitative results, and better signal stability over time.

Classifying and understanding genetic variation between populations and individuals is an important aim in the field of genomics. High resolution melt (HRM) analysis is the quantitative analysis of the melt curve of a DNA fragment following amplification by PCR and can be considered the next-generation application of amplicon melting analysis. Careful sample preparation and planning of experimental and assay design are crucial for robust and reproducible results.

As a fundamental tool in protein research, it is critical that all steps of the electrophoresis workflow (from running gels to visualizing blots) are performed successfully to ensure accurate, reliable, and reproducible results. Protein standards are often used to monitor electrophoretic separation and transfer efficiency but do not always enable ladder visualization after blot development.