merck millipore,默克密理博,S7840,CpG WIZ Oct-4

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merck millipore,默克密理博,S7840,CpG WIZ Oct-4
产品名称：CpG WIZ Oct-4
产品型号：S7840
Primers contained in the CpG WIZ Oct-4 Amplification Kit are designed to specifically amplify each of the sequences based upon these chemically-induced differences.

merck millipore,默克密理博,S7840,CpG WIZ Oct-4

产品介绍

merck millipore,默克密理博,S7840,CpG WIZ Oct-4

描述
产品目录编号	S7840
商名	Chemicon CpG Wiz
描述	CpG WIZ® Oct-4
概述	Principles of the Technique Methylation-specific PCR (MSP), performed using the CpGenome™ DNA Modification Kit and the CpG WIZ® Oct-4 Amplification Kit, permits sensitive detection of altered DNA. Because this is a PCR-based assay, it is extremely sensitive, facilitating the detection of low numbers of methylated alleles and the study of samples containing small amounts of DNA. MSP also allows examination of all CpG sites, not just those with Oct-4 sequences recognized by methylation sensitive restriction enzymes. Increasing the number of such sites that can be assessed allows rapid, fine mapping of methylation patterns throughout CpG regions. In addition, the bisulfite modification is ideally suited for analysis of CpG islands since it converts the majority of cytosines to uracils, making a region of the genome that is CG-rich less difficult to amplify by PCR. MSP employs an initial bisulfite reaction to modify the DNA, followed by a "hot start" PCR amplification with specific primers designed to distinguish methylated DNA from unmethylated DNA. As shown in Figure 1, in the bisulfite reaction, all unmethylated cytosines are converted to uracils while 5-methylcytosines remain unaltered. Thus, the sequence of the treated DNA will differ if the DNA is originally methylated vs. unmethylated. Primers contained in the CpG WIZ® Oct-4 Amplification Kit are designed to specifically amplify each of the sequences based upon these chemically-induced differences. If the sample DNA was originally unmethylated, a product will be generated after PCR using the U primer set. Conversely, a product will be generated using the M primer set if the sample was originally methylated.
背景信息	Oct-4 (POU5F1, Oct-3) is a member of the POU domain family of homeodomain transcription factors, named for its ability to bind octamer DNA binding sites resembling 5'-ATGCAAAT-3'. In mouse, the expression pattern of Oct-4 mRNA is restricted to cells of the inner cell mass in pre- and post-implantation embryos, and primordial germ cells of adult mice (1). Oct-4 expression is essential for maintenance of pluripotentiality of murine ES cells in culture, and withdrawal of Oct-4 activity results in trans-differentiation of embryonic stem cells into trophoectoderm both in culture and in animals (2, 3). As such, Oct-4 represents a well known candidate for a master regulator of stem cell identity, self renewal and maintenance of the undifferentiated state (4). One of the mechanisms implicated in regulation of Oct-4 expression in both mouse and human cells is epigenetic silencing via methylation (5, 6). There is evidence for regulation both at the level of DNA methylation of the Oct-4 regulatory region, as well as chromatin remodeling via methylation of key residues on histone proteins. With respect to DNA, methylation of cytosines located 5' to guanosine is known to have a profound effect on the expression of several eukaryotic genes (7). 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 (8) and has been associated with transcriptional inactivation in various genes (9). Similarly, normal development processes can be controlled by epigenetic inactivation via DNA methylation (imprinting) or promoter methylation of key developmental regulatory genes. Previously developed methods to determine the methylation status of cytosine include digestion with methylation sensitive restriction enzymes and genomic DNA sequencing. Both techniques have limitations: restriction enzymes can only detect methylation sites within their recognition sequence and sequencing is time consuming. Methylation-specific PCR (MSP) is a new technology for sensitive detection of abnormal gene methylation utilizing small amounts of DNA (9). 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. The CpG WIZ® Oct-4 Amplification Kit provides a method for evaluating methylation of the murine Oct-4 promoter in methylation studies (10). Furthermore, this assay provides a sensitive molecular tool to evaluate the differentiation status of murine ES cells in culture. The CpGenome™ DNA Modification Kit (Cat. No. S7820 or S7824) contains the reagents necessary to perform the initial bisulfite reactions, while the CpG WIZ® Oct-4 Amplification Kit contains the reagents required for the PCR amplification reactions.

产品信息
部件	Universal 10X PCR Buffer / 265 μL / 90396 / -15°C to -25°C The components of the CpG WIZ® Oct-4 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 / 2003416 /-15°C to -25°C M Primer Set7.5 μM each primer (25X) / 35 μL / 2003417 / -15°C to -25°C W Primer Set7.5 μM each primer (25X) / 35 μL / 2003511 / -15°C to -25°C

产品信息

部件

Universal 10X PCR Buffer / 265 μL / 90396 / -15°C to -25°C
The components of the CpG WIZ® Oct-4 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 / 2003416 /-15°C to -25°C
M Primer Set7.5 μM each primer (25X) / 35 μL / 2003417 / -15°C to -25°C
W Primer Set7.5 μM each primer (25X) / 35 μL / 2003511 / -15°C to -25°C

应用
应用	Primers contained in the CpG WIZ Oct-4 Amplification Kit are designed to specifically amplify each of the sequences based upon these chemically-induced differences.

生物信息
品种反应性	Mouse
Entrez基因编号	NM_203289.3 NM_002701.4
基因符号	OTF3 POU5F1 Oct-3 OTF4 OCT3 OCT4 Oct-4 MGC22487 Oct4
UniProt编号	Q01860
UniProt汇总	FUNCTION: SwissProt: Q01860 # Transcription factor that binds to the octamer motif (5'-ATTTGCAT-3'). Prime candidate for an early developmental control gene. SIZE: 360 amino acids; 38571 Da SUBCELLULAR LOCATION: Nucleus. TISSUE SPECIFICITY: Ubiquitous. Expressed at low levels. SIMILARITY: SwissProt: Q01860 ## Belongs to the POU transcription factor family. Class- 5 subfamily. & Contains 1 homeobox DNA-binding domain. & Contains 1 POU-specific domain. MISCELLANEOUS: Several pseudogenes of POU5F1 have been described on chromosomes 1, 3, 8, 10 and 12. 2 of them, localized in chromosomes 8 and 10, are transcribed in cancer tissues but not in normal ones and may be involved in the regulation of POU5F1 gene activity in carcinogenesis.

产品使用声明
使用声明	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.