一种检测蛋白酪氨酸激酶的酶联免疫吸附法的描述。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry Pub Date : 1991-02-01 DOI:10.1016/0003-2697(91)90533-Y

Iciar Lázaro, Miguel Gonzalez, Garbiñe Roy, Luisa M. Villar, Pedro Gonzalez-Porqué

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引用次数: 0

摘要

建立了一种检测蛋白酪氨酸激酶的固体免疫分析法。它是基于合成多肽poly(Glu.Na,Tyr) 4:1与微elisa孔的结合，其中磷酸化反应在ATP和酶的存在下发生。最后，用单克隆抗磷酸酪氨酸抗体、过氧化物酶标记的山羊抗小鼠IgG抗体和底物，在同一孔中检测磷酸化酪氨酸残基。样品中蛋白酪氨酸激酶活性的量与每孔中492nm处的颜色成正比。

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Description of an enzyme-linked immunosorbent assay for the detection of protein tyrosine kinase

A solid immunoassay for the detection of protein tyrosine kinases has been developed. It is based on the binding of the synthetic polypeptide poly(Glu·Na, Tyr) 4:1 to microELISA wells, where the phosphorylation reaction takes place in the presence of ATP and enzyme. The phosphorylated tyrosine residues produced in the reaction are finally detected, in the same well, by means of an ELISA using monoclonal antiphosphotyrosine antibody, peroxidase-labeled goat anti-mouse IgG antibody, and substrate. The amount of protein tyrosine kinase activity present in the sample is proportional to the color at 492 nm developed in each well.

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.