A novel binding assay for detection of 14-3-3 protein in cerebrospinal fluid from Creutzfeldt-Jakob patients
| Autor: | Leuck J, Peoch K, Jl, Laplanche, Grabar E, Werner E.G. Müller, Hc, Schröder |
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| Rok vydání: | 2000 |
| Předmět: |
Brain Chemistry
Immunoassay Binding Sites Tyrosine 3-Monooxygenase Amino Acid Motifs Immunoblotting Molecular Sequence Data Phosphoproteins Sensitivity and Specificity Antibodies Creutzfeldt-Jakob Syndrome 14-3-3 Proteins Animals Humans Cattle Amino Acid Sequence Phosphorylation Peptides Aged Protein Binding |
| Zdroj: | Europe PubMed Central |
| ISSN: | 0145-5680 |
| Popis: | Here we describe a simple and fast microplate assay for 14-3-3 protein, which is based on the binding of 14-3-3 protein to a chemically synthesized peptide containing the phosphorylated recognition sequence of this protein. The peptide is covalently bound to maleimide-activated microplates through reaction of the sulfhydryl group of a N-terminal cysteine residue linked to the peptide. The peptide- 14-3-.3 complexes formed are detected using--as a second recognition principle--an anti-14-3-3 protein antibody. This assay was used for measurement of 14-3-3 protein in brain (bovine) and in cerebrospinal fluid from patients with Creutzfeldt-Jakob disease (CJD) and control cases with diseases other than CJD. This novel assay allowed also the detection of low 14-3-3 protein levels present in cerebrospinal fluid, without further concentration, in contrast to conventional immunoblotting or immunoassay procedures. |
| Databáze: | OpenAIRE |
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