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S7815 CpG WIZ® ERα - Methylation specific PCR assay

The components of the CpG WIZ ERalpha Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples. Sufficient reagents are provided to analyze 25 samples.

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S7815
25 assays  
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      Übersicht

      Replacement Information
      Description
      Catalogue NumberS7815
      Brand Family Chemicon®
      Trade Name
      • CpG Wiz
      • Chemicon
      DescriptionCpG WIZ® ERα - 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 (Merlo, 1995). The Estrogen Receptor Alpha (ERalpha) gene is expressed at reduced levels in certain tumor types as a result of hypermethylation of its promoter region (Kim SJ, 2004; Woo IS, 2004).



      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, Cat. No. S7820)
      References
      Product Information
      Components
      • The components of the CpG WIZ® ERalpha 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 Set7.5 μM each primer (25X) / 35 μL (clear cap) / 90633 / -15°C to -25°C
      • M Primer Set7.5 μM each primer (25X) / 35 μL (red cap) / 90634 / -15°C to -25°C
      • W Primer Set7.5 μM each primer (25X) / 35 μL (green cap) / 90635 / -15°C to -25°C
      • U control DNA0.1 μg/μL / 50 μL (clear 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 LevelMQ100
      Applications
      ApplicationThe components of the CpG WIZ ERalpha Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples. Sufficient reagents are provided to analyze 25 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® ERalpha 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 an estrogen receptor, a ligand-activated transcription factor composed of several domains important for hormone binding, DNA binding, and activation of transcription. The protein localizes to the nucleus where it may form a homodimer or a heterodimer with estrogen receptor 2. Estrogen and its receptors are essential for sexual development and reproductive function, but also play a role in other tissues such as bone. Estrogen receptors are also involved in pathological processes including breast cancer, endometrial cancer, and osteoporosis. Alternative splicing results in several transcript variants, which differ in their 5' UTRs and use different promoters.
      Gene Symbol
      • ESR1
      • ESR
      • Era
      • ER
      • NR3A1
      • ESRA
      • DKFZp686N23123
      • ER-alpha
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P03372 # Nuclear hormone receptor. The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues.
      SIZE: 595 amino acids; 66216 Da
      SUBUNIT: Binds DNA as a homodimer. Can form a heterodimer with ESR2. Interacts with NCOA3, NCOA5 and NCOA6 coactivators, leading to a strong increase of transcription of target genes. Interacts with NCOA7 in a ligand-inducible manner. Interacts with PHB2, PELP1 and UBE1C. Interacts with AKAP13. Interacts with CUEDC2. Interacts with JARID1A. Interacts with SMARD1. Interacts with SLC30A9 (By similarity). Interacts with HEXIM1 and MAP1S. Interacts with PBXIP1.
      SUBCELLULAR LOCATION: Nucleus.
      DOMAIN: SwissProt: P03372 Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal steroid-binding domain.
      PTM: Phosphorylated by cyclin A/CDK2. Phosphorylation probably enhances transcriptional activity.
      SIMILARITY: Belongs to the nuclear hormone receptor family. NR3 subfamily. & Contains 1 nuclear receptor DNA-binding domain.
      MISCELLANEOUS: In the absence of ligand, steroid hormone receptors are thought to be weakly associated with nuclear components; hormone binding greatly increases receptor affinity. The hormone- receptor complex appears to recognize discrete DNA sequences upstream of transcriptional start sites.
      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
      Bestellnummer GTIN
      S7815 04053252360978