Antibody probe

Young, R. A., and Davis, R. W. (1983). Efficient isolation of genes using by using antibody probes. Proc. Natl. Acad. Sci USA 80, 1194-1198. 

Fig. 13.3. Fluorescence on the surface of H. contortus intestinal cells following incubation of transverse sections of the worm with fluorescein-labelled antibody probes.

As an alternative, extremely sensitive detection can be achieved with reporter antibody probes tagged with intensely SERS-active compounds or with enzymes that react with substrates to yield SERS-active products. These methods often involve sandwich immunoassay techniques, which increase the number of required steps but offer the advantages of excellent sensitivity and the potential for label multiplexing. For example, Nie and coworkers recently reported the simultaneous detection of two types of antigens in a... 

Proteins bound to the surfaces of synthetic membranes retain their antigenicity and are accessible to antibody probes. The most common membrane-based immunoassay technique is called immunoblotting or, more popularly, Western blotting. In Western blotting, proteins are transferred from an electrophoresis gel to a... 

Burry, R. W., Vandre, D. D., and Hayes, D. M. (1992) Silver enhancement of gold antibody probes in pre-embedding electron microscopic immunocytochemistry. J. Histochem. Cytochem. 40, 1849-1856. 

De Macario, E. C. Macario, A. J. L. Rivard, C. J. Grohmann, K. Use of Antibody Probes to Monitor Hydrolytic Microbes in Municipal Solid Waste Anaerobic Digesters SERI-SP-231-3520 Solar Energy Research Institute Golden, CO, 1989 pp 47-58. 

Fig. 1. Schematic diagram to iiiustrate the principie of western biotting in generating signais from target proteins through antibody probing.

Stripping and re-probing membranes Wash membrane with TBS. Soak membrane in pH2 TBST for 5 min. Rinse the membrane and soak it in pH2 TBST for a further 5 min. Wash membrane with TBST twice. Block membrane and repeat antibody probing (Subheading 3.5). 

Replate the eluted recombinant phage at a density of approx 500-1000 PFU/mL and rescreen with the antibody probe. Repeat the procedure of picking a positive plaque, eluting titrating, and replating at lower plaque densities until it is possible to pick a single well-isolated plaque. 

Young, R. A and Davis, R W (1983) Yeast RNA polymerase II genes isolation with antibody probes. Science 222,778. 

Glenney, J. (1986) Antibody probing of western blots which have been stained with India ink. Anal. Biochem. 156, 315-319... 

Oggero M, Forno G, Kratje R, Etcheverrigaray M (2006), Rational selection of an antibody probe to detect the heterogeneous collection of CHO-derived rhGM-CSF glycoforms, Biotechnol. Lett. 28 2049-2056, DOI 10.1007/sl0529-... 

Acceptable bridging molecule systems have been developed which have also simplified the utilization of different detection systems. To illustrate this point, a researcher who has developed a unique monoclonal antibody (a primary antibody) in the mouse may select from a variety of commercially available products consisting of different detection systems (e.g. fluorescein, alkaline phosphatase, colloidal gold) attached to an immunoglobulin that will specifically bind to mouse antibodies (a secondary antibody). In this way the researcher may readily obtain and test a number of detection methods for visualizing target-probe interactions without having to directly label the monoclonal antibody probe. For nucleic acid probes, which in themselves are not readily immunodetectable, it is useful to incorporate or attach detectable moieties to the nucleotides. 

Biotin has served this purpose well in both nucleic acid and antibody probe systems. As well as being easily detected with immunoglobulins specific for biotin, biotin may also be detected non-immunologically with avidin or streptavidin, two proteins which share a marked, highly specific affinity for biotin. The affinity constant for avidin-biotin interactions is approximately 10 - liters/mole, much higher than the range for antigen-antibody interactions which are commonly between 10 -10 liters/mole. Consequently, a vast number of detection complexes composed of avidin or streptavidin bound to a detection system are commercially available (e.g. streptavidin-alkaline phosphatase). 

The development of monoclonal antibody probes specific to human albumin is the subject of this chapter. These antibody probes and the test methods were developed and evaluated for use in forensic science situations. Although these methods work well on soluble extracts of dried blood stains that are several years old, they have not been applied to archaeological material. The successful development of monoclonal antibodies specific to the human albumin molecule suggests that an approach similar to that used for identifying human tissues and blood on forensic evidence could be applied to any species of interest to the archaeologist. 

Patel N (1994) Imaging neuronal subsets and other cell types in whole-mount Drosophila embryos and larvae using antibody probes. Methods Cell Biol 44 445-187... 

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