Analysis of Suppressor of Cytokine Signalling (SOCS) gene expression by real-time quantitative PCR
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 inhibi...
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Published in | Methods in molecular biology (Clifton, N.J.) Vol. 967; p. 235 |
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Main Authors | , |
Format | Journal Article |
Language | English |
Published |
United States
2013
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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. |
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AbstractList | 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. |
Author | Kolesnik, Tatiana B Nicholson, Sandra E |
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Snippet | 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.... |
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SubjectTerms | 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 |
Title | Analysis of Suppressor of Cytokine Signalling (SOCS) gene expression by real-time quantitative PCR |
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