生物化学技术

Plasmon-waveguide resonance (PWR) spectroscopy is a high-sensitivity optical method for characterizing thin films immobilized onto the outer surface of a glass prism coated with thin films of a metal (e.g., silver) and a dielectric (e.g., silica). Resonance excitation by a polarized cont ...

Fluorescence recovery after photobleaching (FRAP) is a microscopy-based technique that can be used to ask how lipid rafts impact protein and lipid diffusion in cells. This chapter, describes how to perform FRAP measurements of putative raft and nonraft proteins and lipids using a confocal ...

The current models of eukaryotic plasma membrane organization separate the plasma membrane into different environments created by lipids and interactions between membrane proteins and the cytoskeleton, but characterization of their physical properties, such as their size ...

Low-angle X-ray diffraction is a powerful method to analyze the structure of membrane bilayers. Specifically, the technique can be used to determine accurately the thickness of fully hydrated bilayers. Herein details are presented showing how this technique can measure the differen ...

The detection and characterization of lateral heterogeneities or domains in lipid mixtures has attracted considerable interest, because of the roles that such domains may play in biological function. Studies on both model and cell membranes demonstrate that domains can be formed over a ...

The flow of information in cells requires the constant remodeling of cell signaling and trafficking networks. To observe the remodeling events associated with activation of receptors on the cell surface, the authors have generated and analyzed high-resolution topographical maps ...

Atomic force microscopy (AFM) has been a significant tool in the characterization of lipid domains in model membranes. With AFM, one can image the structure of membranes in a natural fluid environment with a lateral resolution that approaches 1 nm and vertical resolution of 0.1 nm. The AFM techniq ...

Computer modeling can provide insights into the existence, structure, size, and thermodynamic stability of localized raft-like regions in membranes. However, the challenges in the construction and simulation of accurate models of heterogeneous membranes are great. The primary ...

Transient lateral microdomains (rafts) in cell membranes have been postulated to perform a number of important functions in normal cells, and are also thought to be critically involved in several pathological conditions. However, there are still a number of fundamental unanswered que ...

The authors solved a microscopic model that describes mixtures of a saturated phospholipid, an unsaturated phospholipid, and cholesterol. The method employed was the self-consistent field approximation. The model was capable of producing several classes of phase diagram, but only ...

Isolation of detergent-resistant membranes (DRMs; also known as detergent-insoluble glycolipid-enriched membranes or glycolipid-enriched membranes ) that are enriched in proteins and lipids with a high affinity for rafts is one of the simplest and most widely used methods for stu ...

Raft and caveolar microdomains have been proposed to participate in numerous cellular functions including signal transduction, cholesterol trafficking, and vesicular sorting. Traditional methods of isolation of rafts from cultured cells and tissue samples have exploited ...

Short-range fluorescence quenching has proven to be an effective method to detect the presence of coexisting ordered and disordered state lipid domains in model membranes. In this approach a fluorescent group and fluorescence-quenching molecule are incorporated into the lipid bi ...

Membrane lipids are structurally diverse in ways that far exceed the role envisioned by Singer and Nicholson of simply providing a fluid bilayer matrix in which proteins reside. Current models of lipid organization in membranes postulate that lipid structural diversity enables nonr ...

Giant unilamellar vesicles (GUVs) are routinely used to study coexisting liquid phases in bilayer membranes. Liquid domains are observed in a wide variety of ternary GUV membranes containing phospholipids and cholesterol, and are thought to model raft domains in cell membranes. GUVs are ...

The integration of immunoassays in microfluidic devices is a rapidly developing research area combining the power of immunoassays with the inherent benefits of microfluidics. Here, a general overview of microfluidic-based immunoassays is presented along with a method for immob ...

The analysis of RNA is central to a wide range of molecular-biology studies, because It 1s often important to obtain informatlon about the expression of genes in hving organlsms Filter hybrldlzatlon of size-separated RNAs with a labeled nucleic-acid probe, a techinque known as northern-bl ...

The detection and identification of specific nucleic acid sequences is routine in molecular biology research The principles that govern molecular annealing (or molecular hybridization) are used in many apphcatlons to the study of nucleic acids. Under sultable conditions, two sin ...

Many important advances in molecular biology would not have been possible wlthout the use of radioisotopes It is relatively simple to substitute a radtoacttve isotope into a nucleotide to produce a molecule that has the same biological properties as the unlabeled molecule (see Chapter 6) T ...

The use of nucleic acid or gene probes as cloning and diagnostic tools has proven to be a powerful technique for lsolatlon and mampulatlon of genes, as well as for rapid detection and ldentlficatlon of bacteria and other mlcroorgamsms, with wide apphcatlons in molecular biology, medical, and env ...