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CpG Wiz? RARb2 Amplification Kit

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    Replacement Information
    Description
    Catalogue NumberS7814
    Brand FamilyChemicon?
    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

    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

    • NM_021976.3

    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

    • P28702

    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
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