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S7809 CpG WIZ® SOCS1 - Methylation specific PCR assay

The components of the CpG WIZ SOCS1 Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples.

S7809
25 assays  
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      Descripción

      Replacement Information
      Description
      Catalogue NumberS7809
      Brand Family Chemicon®
      Trade Name
      • CpG Wiz
      • Chemicon
      DescriptionCpG WIZ® SOCS1 - Methylation specific PCR assay
      OverviewMethylation of cytosines located 5' to guanosine is known to have a profound effect on the expression of several eukaryotic genes (Bird, 1992). In normal cells, methylation occurs predominantly in CG-poor regions, while CG-rich areas, called CpG islands, remain unmethylated. Aberrant methylation of normally unmethylated CpG islands has been documented as a relatively frequent event in immortalized and transformed cells (Antequera, 1990) and has been associated with transcriptional inactivation of defined tumor suppressor genes in human cancers (Herman, 1994; Merlo, 1995). The suppressor of cytokine signaling-1 (SOCS1) gene exhibits characteristic hypermethylation.



      Methylation-specific PCR (MSP) is a new technology for sensitive detection of abnormal gene methylation utilizing small amounts of DNA (Herman, 1996). This process employs an initial bisulfite reaction to modify the DNA, followed by PCR amplification with specific primers designed to distinguish methylated from unmethylated DNA.
      Materials Required but Not DeliveredEquipment and Supplies

      a. Microcentrifuge tubes for PCR amplification

      b. Aerosol-resistant pipette tips

      c. Thermocycler

      d. Gel electrophoresis apparatus (vertical or horizontal)

      e. Power Supply

      f. 302 nm UV transilluminator, camera and film

      Reagents

      a. 2.5 mM dNTP mix (2.5 mM of each nucleotide)

      b. "Hot start" Taq polymerase

      c. "Hot start" PCR reagents (see Sec. II. Protocols).

      d. Reagents for gel electrophoresis (1X TBE and 2% agarose, 10% acrylamide, or suitable high resolution agarose)

      e. DNA markers (size range 100-300 bp)

      f. Ethidium bromide (10 mg/mL)

      g. Gel-loading solution / Loading Dye

      h. Bisulfite Modified DNA (CpGenome™ DNA Modification Kit, S7820)
      References
      Product Information
      Components
      • The components of the CpG WIZ® SOCS1 Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples. Sufficient reagents are provided to analyze 25 samples with appropriate controls.
      • U Primer Set5 μM each primer (25X) 35 μL (neutral cap) 90536 -15°C to -25°C
      • M Primer Set5 μM each primer (25X) 35 μL (red cap) 90537 -15°C to -25°C
      • W Primer Set5 μM each primer (25X) 35 μL (green cap) 90538 -15°C to -25°C
      • U control DNA0.1 μg/μL 50 μL (white cap) 90393 -15°C to -25°C
      • M control DNA0.1 μg/μL 50 μL (red cap) 90394 -15°C to -25°C
      • W control DNA0.05 μg/μL 50 μL (green cap) 90395 -15°C to -25°C
      • Universal 10X PCR Buffer 265 μL (blue cap) 90396 -15°C to -25°C
      Quality LevelMQ300
      Applications
      ApplicationThe components of the CpG WIZ SOCS1 Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples.
      Key Applications
      • PCR
      Application NotesPrinciples of the Technique
      Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® SOCS1 Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.
      Biological Information
      Species Reactivity
      • Human
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a member of the STAT-induced STAT inhibitor (SSI), also known as suppressor of cytokine signaling (SOCS), family. SSI family members are cytokine-inducible negative regulators of cytokine signaling. The expression of this gene can be induced by a subset of cytokines, including IL2, IL3 erythropoietin (EPO), CSF2/GM-CSF, and interferon (IFN)-gamma. The protein encoded by this gene functions downstream of cytokine receptors, and takes part in a negative feedback loop to attenuate cytokine signaling. Knockout studies in mice suggested the role of this gene as a modulator of IFN-gamma action, which is required for normal postnatal growth and survival.
      Gene Symbol
      • SOCS1
      • SSI-1
      • Cish1
      • CISH1
      • SSI1
      • CIS1
      • JAB
      • TIP-3
      • TIP3
      • SOCS-1
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: O15524 # SOCS family proteins form part of a classical negative feedback system that regulates cytokine signal transduction. SOCS1 is involved in negative regulation of cytokines that signal through the JAK/STAT3 pathway. Through binding to JAKs, inhibits their kinase activity. In vitro, also suppresses Tec protein- tyrosine activity. Appears to be a major regulator of signaling by interleukin 6 (IL6) and leukemia inhibitory factor (LIF). Regulates interferon-gamma mediated sensory neuron survival (By similarity). Probable substrate-recognition component of a SCF- like ECS (Elongin-Cullin-SOCS-box protein) E3 ubiquitin ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins. Seems to recognize JAK2.
      SIZE: 211 amino acids; 23551 Da
      SUBUNIT: Interacts with multiple activated signaling proteins of the tyrosine kinase signaling pathway including JAK family kinases, TEC, KIT, GRB2 and VAV. Binding to JAKs is mediated through the KIR and SH2 domains to a phosphorylated tyrosine residue within the JAK JH1 domain. Binds the SH3 domain of GRB2 via diproline determinants in the N-terminus, and the N-terminal regulatory domain of VAV (By similarity). Interacts with the Elongin BC complex (TCEB1 and TCEB2). Component of an ECS CBC(VHL) E3 ubiquitin ligase complex which contains Elongin BC complex, CUL5, RBX1 and SOCS1 (By similarity). Interacts with TRIM8 (By similarity). Interacts with CUL2.
      TISSUE SPECIFICITY: Expressed in all tissues with high expression in spleen, small intestine and peripheral blood leukocytes.
      DOMAIN: SwissProt: O15524 The ESS and SH2 domains are required for JAK phosphotyrosine binding. Further interaction with the KIR domain is necessary for signal and kinase inhibition. & The SOCS box domain mediates the interaction with the Elongin BC complex, an adapter module in different E3 ubiquitin ligase complexes. The Elongin BC complex binding domain is also known as BC-box with the consensus [APST]-L-x(3)-C-x(3)-[AILV] and is part of the SOCS box (By similarity).
      SIMILARITY: Contains 1 SH2 domain. & Contains 1 SOCS box domain.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Packaging Information
      Material Size25 assays
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Número de referencia GTIN
      S7809 04053252616969

      Documentation