| Description | |
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| Catalogue Number | S7808 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | CpG WIZ® GST-pi Amplification Kit |
| 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 (Herman, 1994; Merlo, 1995). Glutathione S-Transferase (GSTpi) is an example of a gene that 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 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, S7820) |
| Product Information | |
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| Components |
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| Quality Level | MQ100 |
| Applications | |
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| Application | The components of the CpG WIZ GSTpi Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples. Sufficient reagents are provided to analyze 25 samples. |
| Key Applications |
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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® GSTpi Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions. |
| Biological Information | |
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| Species Reactivity |
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| Entrez Gene Number |
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| Entrez Gene Summary | Glutathione S-transferases (GSTs) are a family of enzymes that play an important role in detoxification by catalyzing the conjugation of many hydrophobic and electrophilic compounds with reduced glutathione. Based on their biochemical, immunologic, and structural properties, the soluble GSTs are categorized into 4 main classes: alpha, mu, pi, and theta. The glutathione S-transferase pi gene (GSTP1) is a polymorphic gene encoding active, functionally different GSTP1 variant proteins that are thought to function in xenobiotic metabolism and play a role in susceptibility to cancer, and other diseases. |
| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P09211 # Conjugation of reduced glutathione to a wide number of exogenous and endogenous hydrophobic electrophiles. SIZE: 210 amino acids; 23356 Da SUBUNIT: Homodimer. SIMILARITY: SwissProt: P09211 ## Belongs to the GST superfamily. Pi family. |
| Product Usage Statements | |
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| Packaging Information | |
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| Material Size | 25 assays |