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S7814 CpG WIZ® RARβ2 - Methylation specific PCR assay

The components of the CpG WIZ RARbeta2 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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S7814
25 assays  
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      Overview

      Replacement Information
      Description
      Catalogue NumberS7814
      Brand Family Chemicon®
      Trade Name
      • CpG Wiz
      • Chemicon
      DescriptionCpG WIZ® RARβ2 - 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 Retinoic Acid Receptor beta2 is expressed at reduced levels in many tumor types as a result of hypermethylation of its promoter region.



      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® RARbeta2 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) 90595 -15°C to -25°C
      • M Primer Set7.5 μM each primer (25X) 35 μL (red cap) 90596 -15°C to -25°C
      • W Primer Set7.5 μM each primer (25X) 35 μL (green cap) 90597 -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 RARbeta2 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® RARbeta2 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 retinoid X receptor (RXR) family of nuclear receptors which are involved in mediating the effects of retinoic acid (RA). This receptor forms homodimers with the retinoic acid, thyroid hormone, and vitamin D receptors, increasing both DNA binding and transcriptional function on their respective response elements. The gene lies within the major histocompatibility complex (MHC) class II region on chromosome 6. An alternatively spliced transcript variant has been described, but its full length sequence has not been determined.
      Gene Symbol
      • RXRB
      • H-2RIIBP
      • NR2B2
      • MGC1831
      • RCoR-1
      • DAUDI6
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P28702 # Nuclear hormone receptor. Involved in the retinoic acid response pathway. Binds 9-cis retinoic acid (9C-RA).
      SIZE: 533 amino acids; 56922 Da
      SUBCELLULAR LOCATION: Nucleus.
      TISSUE SPECIFICITY: Expressed in a variety of tumor cell lines.
      DOMAIN: SwissProt: P28702 Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal steroid-binding domain.
      SIMILARITY: Belongs to the nuclear hormone receptor family. NR2 subfamily. & Contains 1 nuclear receptor DNA-binding 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
      Catalogue Number GTIN
      S7814 04053252316043

      Documentation

      References

      Reference overviewPub Med ID
      Quantitative RARbeta2 hypermethylation: a promising prostate cancer marker.
      Jerónimo, Carmen, et al.
      Clin. Cancer Res., 10: 4010-4 (2004)  2004

      Show Abstract       		15217932 15217932