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Title: Quantitative determination of 2'-deoxycytidine-5'-triphosphate in cell extracts by radioimmunoassay

Abstract

A radioimmunoassay (RIA) capable of quantitating dCTP in femtomolar amounts in cell extracts has been developed, and applied to human fibroblast cells lines and L5178Y mouse lymphoma lines. Cross reactivity of the antibody with CTP, though low (2.7%) has necessitated pre-RIA removal of CTP by either boronate affinity gel chromatography or sodium periodate oxidation. Fractions from the boronate gel column or aliquots of NaIO/sub 4/-treated cell extract are quantitated directly by the RIA. Recovery of extracted dCTP standard taken through the entire procedure is quantitative and results are reproductive. Due to the high sensitivity of the quantitation step, dCTP can be accurately measured in relatively small numbers of cells-about 10/sup 4/ cells.

Authors:
; ;
Publication Date:
Research Org.:
Univ. of Surrey, England
OSTI Identifier:
5578864
Resource Type:
Journal Article
Journal Name:
Anal. Biochem.; (United States)
Additional Journal Information:
Journal Volume: 154:1; Journal Issue: 1
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; NUCLEOTIDES; RADIOIMMUNOASSAY; ANIMAL CELLS; ANTIBODIES; CHROMATOGRAPHY; FIBROBLASTS; LYMPHOMAS; MICE; OXIDATION; PERIODATES; QUANTITATIVE CHEMICAL ANALYSIS; SENSITIVITY; SODIUM COMPOUNDS; ALKALI METAL COMPOUNDS; ANIMALS; CHEMICAL ANALYSIS; CHEMICAL REACTIONS; CONNECTIVE TISSUE CELLS; DISEASES; HALOGEN COMPOUNDS; IMMUNOASSAY; IMMUNOLOGY; IODINE COMPOUNDS; ISOTOPE APPLICATIONS; MAMMALS; NEOPLASMS; ORGANIC COMPOUNDS; OXYGEN COMPOUNDS; RADIOASSAY; RADIOIMMUNOLOGY; RODENTS; SEPARATION PROCESSES; SOMATIC CELLS; TRACER TECHNIQUES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Piall, E M, Aherne, G W, and Marks, V. Quantitative determination of 2'-deoxycytidine-5'-triphosphate in cell extracts by radioimmunoassay. United States: N. p., 1986. Web. doi:10.1016/0003-2697(86)90526-9.
Piall, E M, Aherne, G W, & Marks, V. Quantitative determination of 2'-deoxycytidine-5'-triphosphate in cell extracts by radioimmunoassay. United States. doi:10.1016/0003-2697(86)90526-9.
Piall, E M, Aherne, G W, and Marks, V. Tue . "Quantitative determination of 2'-deoxycytidine-5'-triphosphate in cell extracts by radioimmunoassay". United States. doi:10.1016/0003-2697(86)90526-9.
@article{osti_5578864,
title = {Quantitative determination of 2'-deoxycytidine-5'-triphosphate in cell extracts by radioimmunoassay},
author = {Piall, E M and Aherne, G W and Marks, V},
abstractNote = {A radioimmunoassay (RIA) capable of quantitating dCTP in femtomolar amounts in cell extracts has been developed, and applied to human fibroblast cells lines and L5178Y mouse lymphoma lines. Cross reactivity of the antibody with CTP, though low (2.7%) has necessitated pre-RIA removal of CTP by either boronate affinity gel chromatography or sodium periodate oxidation. Fractions from the boronate gel column or aliquots of NaIO/sub 4/-treated cell extract are quantitated directly by the RIA. Recovery of extracted dCTP standard taken through the entire procedure is quantitative and results are reproductive. Due to the high sensitivity of the quantitation step, dCTP can be accurately measured in relatively small numbers of cells-about 10/sup 4/ cells.},
doi = {10.1016/0003-2697(86)90526-9},
journal = {Anal. Biochem.; (United States)},
number = 1,
volume = 154:1,
place = {United States},
year = {1986},
month = {4}
}