Please use this identifier to cite or link to this item:
http://hdl.handle.net/1942/3449
Title: | Quantification of cytokine messenger RNA in transfected human T cells by RT-PCR and an automated electrochemiluminescence-based post-PCR detection system | Authors: | MOTMANS, KRIS RAUS, Jef VANDEVYVER, CAROLINE |
Issue Date: | 1996 | Publisher: | ELSEVIER SCIENCE BV | Source: | JOURNAL OF IMMUNOLOGICAL METHODS, 190(1). p. 107-116 | Abstract: | A fast method is reported for the precise and accurate quantification of cytokine mRNA, based on a quantitative polymerase chain reaction (PCR) assay. Post-PCR detection of the amplification products is achieved using an automated electrochemiluminescent (ECL) detection system. The target is amplified using a biotinylated forward and a tris(2,2'-bipyridine)ruthenium (II) (TBR)-labeled reverse primer. The amplification products are then captured on streptavidin coated paramagnetic beads and quantified by measuring the ECL signal of the TBR label. The results obtained are reproducible and accurate over a wide range (3 orders of magnitude) of concentrations. Quantitative results can be obtained using a standard curve which is generated with a synthetic external standard. This technique was applied to quantify tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) mRNA levels in human T cells transfected with the corresponding genes. | Notes: | LIMBURGS UNIV CENTRUM,B-3590 DIEPENBEEK,BELGIUM.Motmans, K, DR L WILLEMS INST,GEHOUW A,UNIV CAMPUS,B-3590 DIEPENBEEK,BELGIUM. | Keywords: | cytokine; mRNA quantification; polymerase chain reaction; tumor necrosis factor-alpha; IFN-gamma; IL-2 | Document URI: | http://hdl.handle.net/1942/3449 | DOI: | 10.1016/0022-1759(95)00259-6 | ISI #: | A1996UD44900011 | Type: | Journal Contribution |
Appears in Collections: | Research publications |
Show full item record
SCOPUSTM
Citations
33
checked on Sep 2, 2020
WEB OF SCIENCETM
Citations
32
checked on Sep 28, 2024
Page view(s)
54
checked on Sep 7, 2022
Google ScholarTM
Check
Altmetric
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.