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Methylated DNA Immunoprecipitation and Microarray-Based Analysis: Detection of DNA Methylation in Breast Cancer Cell Lines

The methylated DNA immunoprecipitation microarray (MeDIP-chip) is a genome-wide, high-resolution approach to detect DNA methylation in whole genome or CpG (cytosine base followed by a guanine base) islands. The method utilizes anti-methylcytosine antibody to immunoprecipi ...

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Use of Reporter Genes to Study the Activity of Promoters in Ovarian Granulosa Cells

Use of reporter genes provides a convenient way to study the activity and regulation of promoters and examine the rate and control of gene transcription. Many reporter genes and transfection methods can be efficiently used for this purpose. To investigate gene regulation and signaling pat ...

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Use of Reporter Genes to Study Promoters of the Androgen Receptor

As a transcriptional regulator, the androgen receptor (AR) regulates the expression of many genes that are essential for male sexual differentiation, including the development of both normal prostate and prostate cancer. The AR acts by binding to regulatory DNA sequences found on the pro ...

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Isolation of Proteins Associated with the DNA-Bound Estrogen Receptor

Regulating gene expression is a complex process requiring the interaction of multiple transcription factors with their cognate recognition sequences. While these DNA-bound transcription factors are the primary drivers of gene expression, the capacity of a transcription fac ...

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Identification of Natural Human Glucocorticoid Receptor (hGR) Mutations or Polymorphisms and Their Functional Consequences at the HormoneReceptor Inte

Glucocorticoids regulate a broad spectrum of physiologic functions essential for life and play an important role in the maintenance of basal and stress-related homeostasis. At the cellular level, the actions of glucocorticoids are mediated by the human glucocorticoid receptor α (h ...

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Genomics Analysis: Endometrium

Microarray technology has been used widely in gynecology. Numerous studies have used this method to address biological questions related to human endometrium. The cyclic changes of endometrium confer special characteristics that should be considered before genomic analysis. ...

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Adenoviral Gene Transfer into Isolated Pancreatic Islets

The beta cells within the pancreatic islets are responsible for production of insulin, a peptide hormone required for maintaining normoglycemia. The establishment of efficient gene transfer into pancreatic islets is very important for studies of insulin and glucagon production a ...

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Detection of Ovarian Matrix Metalloproteinase mRNAs by In Situ Hybridization

In situ hybridization represents a powerful technique to localize DNA or RNA of interest at the chromosomal or cellular level. In endocrine tissues composed of diverse and varied cell types, in situ hybridization has allowed the identification of specific cells responsible for the expre ...

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Basic Molecular Techniques for the Detection of Single Nucleotide Polymorphisms: Genome-Wide Applications in Search for Endocrine Tumor Related Genes

The necessity of genotyping high number of variations in extended sample sets has become apparent in the era of large genomic studies of common complex disorders, in cancer and in pharmacogenomics. The single nucleotide polymorphisms’ (SNPs) apparent advantages over other genetic mar ...

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Breast Tumor-Initiating Cells Isolated from Patient Core Biopsies for Study of Hormone Action

In recent years, evidence has emerged supporting the hypothesis that cancer is a stem cell disease. The cancer stem cell field was led by the discovery of leukemia stem cells (Tan, B.T., Park, C.Y., Ailles, L.E., and Weissman, I.L. (2006) The cancer stem cell hypothesis: a work in progress. Laboratory Investi ...

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Transgenic Mouse Technology: Principles and Methods

Introduction of foreign DNA into the mouse germ line is considered a major technical advancement in the fields of developmental biology and genetics. This technology now referred to as transgenic mouse technology has revolutionized virtually all fields of biology and provided new gen ...

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Real-Time Non-invasive Imaging of ES Cell-Derived Insulin Producing Cells

ES cells are a potential source for insulin producing cells (IPCs). However, two major handicaps are establishing reliable differentiation protocols and the lack of imaging techniques that allow monitoring of these cells post-transplantation. Here, we describe a new approach for mon ...

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Reporter Mice for the Study of Intracellular Receptor Activity

During the past decade the remarkable progress in molecular genetics and the possibility to engineer cells to express genes reporting on the activity of specific promoters has produced major changes in biological research. The description and validation of reporter mice for non-inva ...

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Determining the Affinity of HormoneReceptor Interaction

Characterization of the binding of a hormone to its cognate receptor is a cornerstone of many studies in molecular and cellular endocrinology since this event represents the beginning of a specific cellular response, generally from a highly regulated extracellular messenger. The pre ...

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Use of Laser Capture Microdissection in Studying Hormone-Dependent Diseases: Endometriosis

Endometriosis, a common gynecological disorder responsible for infertility and pelvic pain, is defined as the presence of endometrial glands and stroma within extra-uterine sites. Gene expression studies performed on endometriotic tissue homogenates have yielded results r ...

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Identification of Alternative Transcripts Using Rapid Amplification of cDNA Ends (RACE)

Many organisms, including humans, have many more proteins than are actually coded for by their genes. This discrepancy is partially explained by the existence of alternative transcripts produced by the same gene. Multiple isoforms of the same gene sometimes perform completely differe ...

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Detection of Proteins Sumoylated In Vivo and In Vitro

Small ubiquitin-related modifier (SUMO) is an ubiquitin-like protein that is covalently attached to a variety of target proteins. Unlike ubiquitination, sumoylation does not target proteins for proteolytic breakdown, but is instead involved in regulating multiple protein fun ...

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Detection of ER-SRC-1 Interactions Using Bioluminescent Resonance Energy Transfer

Bioluminescent Resonance Energy Transfer is a naturally occurring phenomenon that can be exploited to explore protein−protein interactions in real-time in intact cells and cellular extracts. It detects energy transferred between a bioluminescent donor enzyme (Renilla luc ...

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Genome-Wide Analysis for ProteinDNA Interaction: ChIP-Chip

Chromatin immunoprecipitation (ChIP) is a well-established procedure for protein−DNA interaction research. ChIP-chip, combining chromatin immunoprecipitation (ChIP) and microarray technology (Chip), enables scientists to survey genome-wide DNA binding sites for a g ...

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Chromosome-Wide Analysis of Protein Binding and Modifications

In order to fully understand the functions of a DNA-binding protein it is necessary to identify all of its binding sites in chromosomes and assess the role of each site in the overall biological function of the factor. An approach ChIP-on-Chip which combines the chromatin immunoprecipitation t ...

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