Back to Search
Start Over
Analysis of Suppressor of Cytokine Signalling (SOCS) gene expression by real-time quantitative PCR.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2013; Vol. 967, pp. 235-48. - Publication Year :
- 2013
-
Abstract
- 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.
- Subjects :
- Bone Marrow Cells cytology
Bone Marrow Cells drug effects
DNA Primers genetics
DNA, Complementary biosynthesis
Humans
Interleukin-4 pharmacology
Macrophages metabolism
Real-Time Polymerase Chain Reaction standards
Reference Standards
Gene Expression Regulation
Real-Time Polymerase Chain Reaction methods
Suppressor of Cytokine Signaling Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 967
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 23296734
- Full Text :
- https://doi.org/10.1007/978-1-62703-242-1_17