Analysis of Suppressor of Cytokine Signalling (SOCS) gene expression by real-time quantitative PCR

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 967; p. 235
• Main Authors: Kolesnik, Tatiana B, Nicholson, Sandra E
• Format: Journal Article
• Language: English
• Published: United States 2013
• Subjects: Bone Marrow Cells - cytology; Bone Marrow Cells - drug effects; DNA Primers - genetics; DNA, Complementary - biosynthesis; Gene Expression Regulation; Humans; Interleukin-4 - pharmacology; Macrophages - metabolism; Real-Time Polymerase Chain Reaction - methods; Real-Time Polymerase Chain Reaction - standards; Reference Standards; Suppressor of Cytokine Signaling Proteins - genetics
• ISSN: 1940-6029
• DOI: 10.1007/978-1-62703-242-1_17

The Suppressor of Cytokine Signalling (SOCS) proteins are a family of negative regulators characterized by a central SH2 domain and C-terminal SOCS box motif. Cytokine Inducible SH2-containing protein (CIS), SOCS1, 2 and 3 are rapidly upregulated in response to cytokine stimulation and act to inhibit JAK/STAT signalling by a variety of mechanisms. The expression of SOCS proteins provides a level of specificity in the control of signalling, with SOCS proteins differentially upregulated in response to individual cytokines and in various cell-types. Real-time reverse transcription (RT) quantitative polymerase chain reaction (RT-qPCR) is an established technique for quantifying mRNA in biological samples, measuring the relative expression of genes of interest and identifying single nucleotide polymorphisms. Here we describe the use of SYBR(®) Green I RT-qPCR to quantify the relative expression level of SOCS mRNA in murine bone marrow-derived macrophages (BMDM). The approach can be universally applied to different cell types and various tissues.