Detection system protein

Most commercial protein detection systems in use today are based on the specific recognition of antigens with antibodies in several immunoassay formats [43]. 

Takahashi, M. Nokihara, K. Mihara, H., Construction of a protein-detection system using a loop peptide library with a fluorescence label. Chem. Biol. 2003, 10, 53-60... 

Jenkins, D. B. Tucker, E. B. Kozikoski, T. E. Protein detection system in mammalian urine using phloxine... 

While the fluid mosaic model of membrane stmcture has stood up well to detailed scrutiny, additional features of membrane structure and function are constantly emerging. Two structures of particular current interest, located in surface membranes, are tipid rafts and caveolae. The former are dynamic areas of the exo-plasmic leaflet of the lipid bilayer enriched in cholesterol and sphingolipids they are involved in signal transduction and possibly other processes. Caveolae may derive from lipid rafts. Many if not all of them contain the protein caveolin-1, which may be involved in their formation from rafts. Caveolae are observable by electron microscopy as flask-shaped indentations of the cell membrane. Proteins detected in caveolae include various components of the signal-transduction system (eg, the insutin receptor and some G proteins), the folate receptor, and endothetial nitric oxide synthase (eNOS). Caveolae and lipid rafts are active areas of research, and ideas concerning them and their possible roles in various diseases are rapidly evolving. 

Light scattering (nephelometry) was used as a detection system for gly-cosaminoglycans from urine, eluted from a DEAE Sephadex (Pharmacia Biotechnology Uppsala, Sweden) A-25 column.68 This technique has been more recently applied to protein characterization.69 Interferometry was used for analysis of dextran eluted from a size exclusion column.70 One of the problems of electrochemical detection is that it is relatively insensitive to polymers. Because many of the materials discussed below (DNA, proteins, and polysaccharides) are polymeric, a brief mention of some alternative... 

B. Schmidt and D. Riesner, A fluorescence detection system for the analytical ultracentrifuge and its application to proteins, nucleic acids, viroids and viruses (in Ref. [77]). 

The presence of biotin labels on an antibody molecule provides multiple sites for the binding of avidin or streptavidin. If the biotin binding protein is in turn labeled with an enzyme, fluoro-phore, etc., then a very sensitive detection system is created. The potential for more than one labeled (strept)avidin to become attached to each antibody through multiple biotinylation sites provides an increase in detectability over antibodies directly labeled with a detectable tag. 

This section discusses the properties of gold particles as well as the common methods of labeling proteins and other biomolecules with them. The cited references should be consulted to obtain protocols for using these protein-gold complexes in assay and detection systems. 

H. Ge, UPA, a universal protein array system for quantitative detection of protein-protein, protein-DNA, protein-RNA and protein-ligand interactions. Nucl. Acids Res. 28, e3 (2000). 

In any chromatographic analysis the method of detection is determined by the nature of the analyte and the mobile phase used must not interfere with this system. The use of ultraviolet absorption detection systems is very common but the solvents used must not absorb significantly at the wavelength used. For instance, absorption at 280 nm is frequently used to detect protein but some solvents, e.g. acetone, absorb at this wavelength. Similarly the use of concentration gradients in the mobile phase may present problems with refractive index and electrochemical detection systems. 

Gel electrophoresis provides a simple method for separating complex protein mixtures. Because proteins are visualized using stains that may not be linearly incorporated in the gel, the intensity of the stained bands may be poorly correlated with the amount of protein. For this reason, gel electrophoresis is at best a semiquantitative technique capable of generating relative purity results. In CE, separations are commonly performed in free solution, i.e., in the absence of any support such as gel matrices. This allows the replacement of the capillary s content in between analyses and therefore the automation of the process. The use of UV-transparent fused-silica capillaries enables direct on-line optical detection of focused protein zones, eliminating the requirement for sample staining. The detection systems available to CE provide true quantitative capabilities. 

Detection system lacks sensitivity necessary to detect the amount of protein loaded... 

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