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A Combination of Affinity Chromatography, 2D DIGE, and Mass Spectrometry to Analyze the Phosphoproteome of Liver Progenitor Cells

Reversible protein phosphorylation is a ubiquitous posttranslational modification that regulates cellular signaling pathways in multiple biological processes. A comprehensive analysis of protein phosphorylation patterns can only be achieved by employing differ ...

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Phosphoproteomic Analysis of Liver Homogenates

Regulation of protein function via reversible phosphorylation is an essential component of cell signaling. Our ability to understand complex phosphorylation networks in the physiological context of a whole organism or tissue remains limited. This is largely due to the technical ch ...

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Glycoproteins and Glycosylation: Apolipoprotein C3 Glycoforms by Top-Down MALDI-TOF Mass Spectrometry

Apolipoprotein C3 (ApoC3) is synthesized in liver so that levels or isoform distributions may constitute indicators of liver pathogenesis. The glycoforms of intact protein ApoC3 in serum or plasma can be readily analyzed by matrix-assisted laser desorption/ionization-time of fl ...

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Cell Surface Capturing Technologies for the Surfaceome Discovery of Hepatocytes

Proteins expressed at the cell surface define how cells can functionally interact with their microenvironment in time and space. The cell surface subproteome, or surfaceome, represents a cellular information gateway not only enabling the processing of environmental molecular c ...

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Plasma Membrane Isolation Using Immobilized Concanavalin A Magnetic Beads

Isolation of highly purified plasma membranes is the key step in constructing the plasma membrane proteome. Traditional plasma membrane isolation method takes advantage of the differential density of organelles. While differential centrifugation methods are sufficient to e ...

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Discovery of Lamin B1 and Vimentin as Circulating Biomarkers for Early Hepatocellular Carcinoma

The recent advancements in proteomic technologies have reconstituted our research strategies over different type of liver diseases including hepatocellular carcinoma (HCC). Combined analyses on HCC proteome and clinicopathological data of patients have allowed identi ...

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Targeted Mass Spectrometry-Based Metabolomic Profiling Through Multiple Reaction Monitoring of Liver and Other Biological Matrices

In a systemic viewpoint, relevant biological information on living systems can be grasped from the study of small, albeit pivotal molecules which constitute the fundamental bricks of metabolic pathways. This holds true for liver which plays, among its unique functions, a key role in metabo ...

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Combinatorial Peptide Ligand Libraries to Discover Liver Disease Biomarkers in Plasma Samples

High-abundance proteins present in blood plasma make the detection of low-abundance proteins extremely difficult by proteomics technology. Hexapeptide combinatorial ligand libraries can be used to investigate the hidden proteome in depth. Here we describe how liver disease bi ...

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An On-Target Desalting and Concentration Sample Preparation Protocol for MALDI-MS and MS/MS Analysis

2DE coupled with MALDI-MS is one of the most widely used and powerful analytic technologies in proteomics study. The MALDI sample preparation method has been developed and optimized towards the combination of simplicity, sample-cleaning, and sample concentration since its introdu ...

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Isolation of Urinary Exosomes from Animal Models to Unravel Noninvasive Disease Biomarkers

In the last years, disease biomarker discovery has highly evolved thanks to the application of high-�throughput technologies such as proteomics. However, due to the elevated complexity and abundance of some of the proteins in the samples the analysis of subcellular compartments has been ...

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Proteomic Methods for Biomarker Discovery in a Rat Model of Alcohol Steatosis

Understanding changes in the expression of specific proteins and/or alterations in their posttranslational modifications is crucial to elucidating the molecular mechanisms underlying disease states such as alcoholic liver disease. Protein separation and analysis tech ...

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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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