2-D electrophoresis followed by immunodetection of protein spots using western blotting is adopted commonly in proteomics studies. Here we present step-by-step instructions for obtaining accurate overlaps between the gel and the blot using the PDQuest 2-D gel analysis software.

Ligand-capture approaches using biotin as an affinity tag are often utilized in biomolecular interaction analyses with surface plasmon resonance (SPR) biosensors. However, a drawback to using biotin is that it makes surface regeneration of SPR sensor chips difficult. Here is a strategy that facilitates surface regeneration when using biotinylated ligand-capture approaches to SPR analysis.

Surface Plasmon Resonance (SPR) is an optical phenomenon that is used for the label-free analysis of the binding of any two molecules in real-time.

Amine coupling is the most commonly adopted technique to bind proteins to biosensor chips. Because of the possibility of the amine coupling reagents reacting with each other, these have to be premixed just before the injection and injected independently. Here are some tips to automatically coinject these reagents.

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.

Protein quantitation is an important part of many workflows. The estimation or quantitation of the protein in a sample frequently allows researchers to understand the results from their work and make decisions on the subsequent steps. Presented are answers to some of the most frequently ask questions related to usage of Bio-Rad's colorimetric assays for protein quantitation.

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.