Protocols and Tips

This appendix to “Guide to SPR Data Analysis on the Proteon™ XPR36 System” gives information on the data set export using ProteOn Manager™ Software.

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.

Learning to use analysis software can be time-consuming and following a manual can be challenging.

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.

Now you can optimize research expenditure by multiplexing only the targets you need to get the answers you’re seeking. This newly updated tool allows you to choose from within panels to build the perfect kit.

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.