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J Immunol Methods. 1994 Dec 28;177(1-2):23-8.

A novel multispecific competitor fragment for quantitative PCR analysis of cytokine gene expression in rats.

Author information

1
Institute for Medical Immunology, Medical Faculty (Charité), Humboldt University, Berlin, Germany.

Abstract

Competitive polymerase chain reaction (PCR) is a sensitive method for quantification of cytokine mRNA expression. Co-amplification of an internal standard serves as control for comparing the efficiency of PCR in different samples. We have developed a novel control fragment for multiple analyses of rat cytokine gene expression containing primers for IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-10, TNF-alpha, TGF-beta 1, IFN-gamma and MIP-2. Additional primers were incorporated to analyse the content of T cells (CD3), activated T cells (CD25) and housekeeping genes (beta-actin and HPRT). As an example we demonstrate analysis of IL-2 mRNA expression in small pieces of kidney tissue obtained from rats after kidney allotransplantation. The IL-2 expression decreased tenfold during treatment with an anti-rat CD4 monoclonal antibody as compared to untreated animals.

PMID:
7822831
[Indexed for MEDLINE]

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