Document Detail


A new electrochemical assay method for gene expression using HeLa cells with a secreted alkaline phosphatase (SEAP) reporter system.
MedLine Citation:
PMID:  22331791     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A new electrochemical assay for the detection of secreted alkaline phosphatase (SEAP) from transfectant HeLa cells is proposed using a microarray device and scanning electrochemical microscopy (SECM). The assay consists of two steps: the first is the incubation of a transfected cell in a microarray culture device covered with a substrate modified with anti-SEAP under physiological conditions without any additives. The array device consists of a 4 × 4 array of microwells having a size of 100 × 100 µm (diameter × depth). The second step is SECM measurement of secreted SEAP at the antibody-immobilized substrate. This assay ensures accuracy and intactness because the undesired influence of endogeneous ALP is eliminated and the transfected cells are incubated in a culture device under suitable conditions. We successfully detected the expression of SEAP from intact cells at the single-cell level using this assay. The system is useful as a cell-based gene-expression assay. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc.
Authors:
Mustafa Sen; Kosuke Ino; Hitoshi Shiku; Tomokazu Matsue
Publication Detail:
Type:  LETTER     Date:  2012-2-13
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  -     ISSN:  1097-0290     ISO Abbreviation:  -     Publication Date:  2012 Feb 
Date Detail:
Created Date:  2012-2-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012 Wiley Periodicals, Inc.
Affiliation:
Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki, Aoba, Sendai 980-8579, Japan.
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