生物化学技术

Along with other resonance energy transfer techniques, bioluminescence resonance energy transfer (BRET) has emerged as an important method for demonstrating protein–protein interactions in cells. In the field of G-protein-coupled receptors, including chemokine recept ...

This chapter describes assays that focus on the characterization of compounds identified in high-�throughput screening campaigns, and the subsequent medicinal chemistry programs. They cover methods to determine potency in buffer, the effect of whole blood on the compounds’ activ ...

Virus-like particles (VLPs) presenting conformational envelope proteins on their surface represent an invaluable tool to study molecular interactions between viruses and cellular receptors/co-receptors, eliminating biological risks associated with working with l ...

Neural stem cells (NSCs) represent a limited population of progenitor cells in the central nervous system that sustain their self-renewal and multipotency from early development to adulthood. Recent evidence suggests that chemokine receptors are constitutively expressed by N ...

Quantifying protein and RNA expression within specific cell populations in vivo is an essential step in unraveling the complex mechanisms of neurological disease. The challenges associated with studying human brain tissue are commonly compounded by variations in postmortem in ...

Chemokines and their G-protein-coupled receptors play important roles in development, homeostasis, and the innate and adaptive immune response. Pathologic chemokine signaling pathways in the peripheral nervous system can be studied in peripheral nerves using human in vitro mo ...

The action of chemokines (or “chemotactic cytokines”) is recognized as an integral part of inflammatory and regulatory processes. Leukocyte mobilization during physiological conditions, trafficking of various cell types during pathological conditions, cell activation, ...

Successful innovative proteomic analysis is highly dependent on molecular biology techniques at the �surveying and validation stage. This is because mass spectrometry (MS) analyses of complex samples are limited by their dynamic range for detection—so careful front-end sample p ...

Leukocyte migration through the interstitial space is crucial for the maintenance of tolerance and immunity. The main cues for leukocyte trafficking are chemokines thought to directionally guide these cells towards their targets. However, model systems that facilitate quanti ...

Fluorescent activated cell sorting (FACS) is a powerful technique that allows rapid quantitative cell-by-cell analysis of the expression of multiple distinct proteins and the isolation of specific cell types from complex cellular mixtures. FACS-based detection of chemokine re ...

Epigenetic regulation of gene expression is an important event for normal cellular homeostasis. Gene expression may be “switched” on or “turned” off via epigenetic means through adjustments in DNA architecture. These structural alterations result from changes to the DNA methylati ...

In response to central nervous system (CNS) injury and infection, astrocytes, neurons, and CNS vasculature express several chemokines, including CCL21. Quantitative polymerase chain reaction (qPCR), western blot, and immunohistochemical methods can quantify mRNA and prote ...

Experimental autoimmune encephalomyelitis (EAE) is a CD4 T cell-mediated demyelinating disease of the central nervous system (CNS) where macrophages are the end-stage effector cell. EAE serves as a model for multiple sclerosis where it has been instructive in delineating the autoim ...

Fibrates are peroxisome proliferator-activated alpha receptor (PPARα) activators derived from fibric acid and are the most clinically used therapeutics in the treatment of hypertriglyceridemia. Recently, we reported a computational approach for the investigation of the bi ...

The capacity to induce the association of peroxisome proliferator-activated receptors (PPARs) with different transcriptional coregulators is determined by the peculiar 3D-structure that the receptors adopt when bound with a specific ligand. The fluorescence resonance en ...

Proteins carry out cellular functions. Identifying proteins within tissues, which are characteristically comprised of various cell types, is critical to understanding how the tissue functions. Being able to assess protein expression in tissues is also essential to gaining insig ...

Peroxisome proliferator-activated receptors (PPARs) are key nuclear receptors which mediate the regulation of the transcription of many genes, especially those involved in lipid catabolism and inflammation. In mammals, three types of PPARs named PPARα, PPARβ (also called PPARδ), ...

Peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) is a nuclear transcriptional coactivator that regulates the genes involved in energy metabolism. Recent evidence has been provided that alternative splicing of PPARGC1A gene produces a functio ...

The role of certain amino acids in the interactions of ligands with their cognate nuclear receptors is usually achieved by the resolution of the crystal structure of the receptor complexed with the ligand. As a complementary functional approach, site-directed mutagenesis, a technique b ...

The nuclear receptor peroxisome proliferator activated receptor gamma (PPARγ) is most abundantly expressed in adipose tissue and has been shown to play imperative roles in controlling adipogenesis and lipogenesis in cultured cell systems in vitro as well as in mice and humans. However, ...