详细说明
| Replacement Information |
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| Description | |
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| Catalogue Number | S7814 |
| Brand Family | Chemicon® |
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| Description | CpG WIZ® RARβ2 - Methylation specific PCR assay |
| Overview | Methylation 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 Delivered | Equipment 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) |
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| Application | 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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| Application Notes | Principles 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 | |
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| Entrez Gene Summary | This 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. |
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| UniProt Summary | FUNCTION: 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. |
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| Material Size | 25 assays |
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