Top Stories of 2012 – BioRadiations Year in Review

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.

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.

Pathway analysis is an efficient approach for studying a large number of related gene targets in a single experiment. Bio-Rad Laboratories and Thomson Reuters, a leading provider of system biology tools, have partnered to provide predesigned pathway panels for gene expression analyses using real-time qPCR. This tech report describes how Bio-Rad’s PrimePCR™ pathway panels were curated and designed using a ranking strategy developed by Thomson Reuters.

When Dr. Fred Bauzon started his research, he worked with bioinformatics specialists to functionalize suitable gene candidates based on their expression levels with HMGCR transcript levels. All his experiments demonstrated a reduction in low-density lipoprotein receptor protein (LDLR) and candidate gene protein levels after the candidate gene was silenced. Here Bauzon discusses how TGX Stain-free precast gels have aided his research toward future drug targets for the therapeutic control of LDL uptake.

The MIQE guidelines have set a new standard for publishing qPCR results to ensure integrity in the scientific literature and increase experimental transparency. One of the key guidelines focuses on the proper design and experimental validation of primer assays used in qPCR. This tech report describes the design and wet-lab validation of PrimePCR assays and how they comply with the MIQE guidelines.

alt Melanoma is an aggressive cancer where early detection is essential for patient survival. The Facchiano lab is developing new anti-proliferation molecules against melanoma and is working to identify new molecular markers at the very early stages of pathology. Here Dr Antonio Facchiano discusses the merits of using the Bio-Plex technology to measure dose-dependent changes in expression levels of growth and angiogenic factors in cell lysates and supernatants.

Sangamo BioSciences, Inc., located in Richmond, California, is developing HIV therapeutics toward a functional cure using their proprietary zinc finger DNA-binding protein technology. Droplet digital PCR (ddPCR™) — or third generation PCR — offers a superior solution as it can detect target sequence within a single molecule, making it a powerful technology for low copy event detection.

Although the Western blot is a core lab procedure, surprisingly little has changed in traditional bench work practice since its inception in 1979. The V3 Western Workflow delivers a streamlined solution to the conventional western blotting workflow as explored here by one protein researcher specializing in chronic obstructive pulmonary disease (COPD).

The T-helper cell (Th17) pathway has been implicated in several autoimmune diseases and in cancer. New magnetic bead–based assays have been developed for several biomarkers in this pathway. The performance characteristics of these assays are evaluated from data on intra- and inter-assay precision, standard curve recovery, cross reactivity, and linearity of dilution.

The significant drop in leprosy worldwide is attributed to the development of antimicrobial multidrug therapy (MDT). Resistance to rifampicin — the backbone of MDT treatment — has appeared. Controlling leprosy transmission requires routine surveillance for mutations in the drug target genes before, during, and after the course of treatment. New high resolution melt (HRM) assays offer leprosy researchers a faster, more economical method to investigate resistance targets.

The McCarroll laboratory studies the biological effects of human genome polymorphism, seeking to define how genome variation influences gene expression and risk of disease.

Bio-Rad has long demonstrated innovation in imaging, with the introduction of the stain-free gel electrophoresis workflow, which was rapidly followed by the launch of the Gel Doc™ EZ imager. Now, this progression continues with the release of the ChemiDoc™ MP imaging system.

Hexapeptide libraries, such as Bio-Rad’s ProteoMiner™ protein enrichment technology, can significantly increase sensitivity of shotgun proteomics analyses of diverse complex samples. Underlying this increased sensitivity is the partial removal of high-abundance proteins leading to an increased representation of low-abundance proteins — a phenomenon described as dynamic range compression. Currently, hexapeptide libraries have been mostly used to increase proteomic detection in fluid samples (eg: saliva, plasma, etc.), but not in cellular lysates.

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.

The genes coding for membrane proteins, such as G-protein coupled receptors (GPCR), ion channels, and other membrane-bound enzymes make up 25–30% of the human genome and membrane proteins are estimated to represent 30–40% of the top drug targets. There is growing research, industrial, and commercial interest in the study of membrane proteins in general, and more specifically in immobilizing membrane proteins to various biosensor surfaces.