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Titolo:
Chemiluminescent standards for quantitive comparison of two-dimensional electrophoresis western blots
Autore:
Conrad, CC; Malakowsky, CA; Talent, J; Rong, D; Lakdawala, S; Gracy, RW;
Indirizzi:
Univ N Texas, Ctr Hlth Sci, Dept Mol Biol & Immunol, Mol Aging Unit, Ft Worth, TX 76107 USA Univ N Texas Ft Worth TX USA 76107 Mol Aging Unit, Ft Worth, TX 76107 USA
Titolo Testata:
PROTEOMICS
fascicolo: 3, volume: 1, anno: 2001,
pagine: 365 - 369
SICI:
1615-9853(200103)1:3<365:CSFQCO>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
SENSITIVE QUANTIFICATION; PROTEINS; GELS;
Keywords:
chemiluminescence; standards; two-dimensional electrophoresis;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
9
Recensione:
Indirizzi per estratti:
Indirizzo: Gracy, RW Univ N Texas, Ctr Hlth Sci, Dept Mol Biol & Immunol, Mol Aging Unit, Ft Worth, TX 76107 USA Univ N Texas Ft Worth TX USA 76107 Unit, Ft Worth, TX 76107 USA
Citazione:
C.C. Conrad et al., "Chemiluminescent standards for quantitive comparison of two-dimensional electrophoresis western blots", PROTEOMICS, 1(3), 2001, pp. 365-369

Abstract

Chemiluminescent probes offer highly sensitive quantitative analyses of proteins blotted from electrophoretic gels onto a supporting matrix (e.g. nitrocellulose or polyvinylidene difluoride). Visualization of signals from probes involves the emission of light that is dependent on a number of variables (e.g. conjugated enzyme activity, antibody titer, hybridization efficiency, substrate concentration, chemical reactivity, temperature, etc.). Thus, it is important to be able to correct for these variations. For example, the exposure time of the blot to the detection medium (e.g., film or digital camera) is a critical variable in the final results. Several protein samples separated on a single blot (e.g. one-dimensional resolution) can be compared from the ratio of the individual proteins, but comparison of separateblots completed on different days requires a chemiluminescent standard. The situation is more complex when only one sample per gel/blot is used (i.e.two-dimensional electrophoresis (2-DE)). This paper describes a method forpreparing agarose embedded standardized strips that contain dilutions of antigens that can be visualized with the corresponding probe antibody. This standardization strip can be produced from common laboratory supplies and provides a method to correct for alterations in chemiluminescent intensitiesfrom different 2-DE analysis. Several standardization strips can be produced simultaneously and then used during the electroblotting step of different blots on different days.

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Documento generato il 21/01/20 alle ore 06:54:15