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丁香实验推荐阅读
Proteomic Study of Caveolae and Rafts Isolated From Human Endothelial Cells

Caveolae and rafts are specialized microdomains of the endothelial cell plasma membrane, which play an important role in signal transduction, transcellular transport, and cholesterol homeostasis. The dynamic protein composition of these subcellular lipid domains has been im ...

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Analysis of RNA by Northern-Blot Hybridization

Northern-blot hybridization, also referred to as Northern blotting, is one of several methods developed to detect the presence, to determine the size, and to quantify specific cellular mRNAs. By this method, total RNA or poly(A)+mRNA, prepared from the cells or tissue of interest, is fractiona ...

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Chipping Away at Heart Failure

Studies in the field of microarray technology have exploded onto the scene to delve into the unknown underlying mechanisms and pathways in molecular disease. Diseases of the cardio-vascular system, particularly those with unexplained molecular etiologies, such as heart failure, h ...

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Isolation and Culture of Human Alveolar Type II Pneumocytes

Alveolar type II pneumocytes (alveolar type II cells; TII cells) play an important role in the homeostasis of the alveolar unit. They are the progenitor cells to the type I pneumocyte and are therefore responsible for regeneration of alveolar epithelium following alveolar epithelial cell ...

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Assay of Endotoxin by Limulus Amebocyte Lysate

Horseshoe crabs fight off infectious agents with a complex array of proteins present in amebocytes, the major cell type in their hemolymph. These amebocytes contain both large and small granules (1). When exposed to bacteria or other infectious agents the amebocytes release proteins into t ...

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Preparation of Endotoxin from Pathogenic Gram-Negative Bacteria

Endotoxins have been recognized for decades as important structural components of the outer cell wall/cell membrane complex of Gram-negative microorganisms. These chemically heterogeneous macromolecular structures were recognized very early on to consist of lipid, polys ...

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Assay of Anti-Endotoxin Antibodies

Lipopolysaccharides (LPS) constitute components of the outer membrane of Gram-negative bacteria. Chemically, they consist of a heteropolysaccharide and a covalently linked lipid, termed lipid A. The polysaccharide region is made up of the O-specific chain (built from repeating u ...

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Isolation of the Bactericidal/Permeability-Increasing Protein from Polymorphonuclear Leukocytes by Reversible Binding to Target Bacteria

Polymorphonuclear leukocytes (PMN) play a prominent role in host defense in mammals against invading bacteria (1,2). Among the essential attributes of these highly specialized cells are the elaboration of an array of cytotoxic peptides and polypeptides that can be targeted at bacteri ...

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Purification of Lipopolysaccharide Binding Protein

Lipopolysaccharide (LPS)-binding protein (LBP) is a critical component of innate immunity, implicated in the initiation of host defences against Gram-negative bacteria. LBP alerts the host to the presence of minute amounts of LPS (1). LPS released from Gram-negative bacteria is prese ...

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Purification of Streptococcal Pyrogenic Exotoxin A

Group A streptococci secrete a variety of molecules, many of which are recognized as virulence factors important in the establishment of streptococcal infections. Among these extracellular products is streptococcal pyrogenic exotoxin A (SPE A, scarlet fever toxin A, erythrogenic ...

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Assay for Superantigens

This chapter describes a quantitative assay for the streptococcal superantigen, streptococcal pyrogenic exotoxin A (SPEA), which can be used in broth, tissue-culture media, and certain sera. The protocol can be adapted to allow measurement of any bacterial superantigen or protein to ...

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Whole-Blood Assays for Cytokine Production

Substantial interest has been generated by the potential roles of the cytokines in health and disease (1). This has prompted considerable investigation into how these mediators are regulated to answer such basic questions as which stimuli initiate transcription and what factors are re ...

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Bioassay for Tumor Necrosis Factors- and

The realization that much of the toxicity of bacterial endotoxin resulted from production of a macrophage-derived intermediate (1) led to the isolation and cloning of tumor necrosis factor (TNF), also known as cachectin (2). Since then, much evidence has accumulated to demonstrate that TNF ...

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Assay of Soluble Tumor Necrosis Factor Receptors

Over the last decade, numerous basic biological as well as experimental and clinical studies have firmly established the significance of tumor necrosis factor (TNF) as a principal proximal mediator of sepsis (1–4). One of the major insights that has emerged during recent years has been that un ...

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Nitric Oxide Synthase Inhibitors

In 1990, several studies reported that an enhanced formation of endogenous nitric oxide (NO) contributes to the hypotension caused by endotoxin and tumor-necrosis factor-α (TNFα) (1–3). In addition, it became apparent that this overproduction of NO also plays an important role in the patho ...

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Blastocyst Culture

Experimental models of blastocyst development based on in vitro culture have played a prominent role in advancing our understanding of peri-implantation development, a process that is relatively inaccessible in vivo. Blastocyst culture provides a robust approach for examining ...

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Isolation of Hormone Responsive Uterine Stromal Cells: An In Vitro Model for Stromal Cell Proliferation and Differentiation

The female sex hormones estrogen and progesterone stimulate proliferation and differentiation of human and rodent uterine cells. The purpose of this chapter is to provide a method for isolating hormone-responsive rat uterine stromal cell lines that can be used to study steroid control of ...

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Rat Decidual Cell Cultures

Pregnancy requires profound reorganization of the different tissues forming the uterus. Growth and differentiation of the uterine endometrial cells give rise to the decidual tissue, a transitory organ, which plays a key role in fetal survival. In this chapter, we describe a technique for t ...

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The Immortalization of Human Endometrial Cells

The loss of replicative potential with each cell division has been attributed to the progressive shortening of telomeres. This “mitotic clock” occurs because most normal human cells are telomerase-negative. Telomerase is a multicomponent enzyme that prevents loss of telomeric DNA a ...

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Sheep Uterine Gland Knockout (UGKO) Model

Endometrial gland development is a postnatal event in the ovine uterus that can be inhibited epigenetically by chronic exposure of ewe lambs to a synthetic progestin after birth. The uterus of neonatally progestinized ewes lack endometrial glands and display a uterine gland knockout (U ...

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