A rapid filtration assay for cAMP.

Abstract:

:The receptor-binding assay for cAMP was improved by using polyethylenimine-treated glass filters. A polyethylenimine-treated glass filter has high protein binding capacity. This high capacity allows an increase in the amount of protein per assay tube and the use of a crude preparation, such as a beef heart extract, as specific binding protein instead of a purified protein, which has been used in the classical filtration assays involving cellulose ester filters. Since the time required for the separation of the protein-cAMP complex and the free nucleotide can be shortened by the use of polyethylenimine-treated filters, the dissociation of the bound ligand during the separation procedure, which is a serious problem with other modified assay methods involving charcoal adsorption, is minimized. Filtration through polyethylenimine-treated glass filters also gives low blanks and prevents the loss of protein or ligand due to breakage of the filters, which is often observed with fragile cellulose ester membranes. In consequence, this simple and rapid filtration assay allows more accurate and reproducible determinations.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Takeda T,Kuno T,Shuntoh H,Tanaka C

doi

10.1093/oxfordjournals.jbchem.a122662

subject

Has Abstract

pub_date

1989-02-01 00:00:00

pages

327-9

issue

2

eissn

0021-924X

issn

1756-2651

journal_volume

105

pub_type

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