Titlebook: Chemokine Protocols; Amanda E. I. Proudfoot,Timothy N. C. Wells,Christi Book 2000 Humana Press 2000

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Chemokine Expression in Insect Cells,tivities of gene products. Thus, efficient expression of recombinant proteins of cloned genes is essential to obtain proteins sufficient for detailed biological and biochemical analysis. In this chapter, I will describe a rapid and efficient method to generate recombinant chemokines in insect cells (.,.).

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Purification of Chemokines from Natural Sources,walking strategy, we have discovered an ever-increasing number of novel genes encoding chemokines that were expressed in bacteria or eukaryotic cells and subsequently tested as the recombinant form for its biological activity.

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Cloning of Novel Chemokines Using a Signal Sequence Trap Method,tively clone cDNA species encoding intercellular signal-transducing molecules without biologic assays. In this chapter, an efficient signal sequence trap method based on an Epstein-Barr virus shuttle vector is described (.).

DEMN

Chemokine Receptor Cloning,to bind chemokines (.). Reverse methods, which involve cloning a cDNA or gene without primary knowledge of the structure of the encoded protein, can be divided into two types: expression and homology hybridization cloning.

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Generation of Stable Cell Lines Expressing Chemokine Receptors, lines expressing high levels of chemokine receptors is also becoming increasingly important in high throughput screening approaches for the identification of chemokine receptor antagonists and agonists. The basic steps in the generation of stable cell lines expressing chemokine receptors are as follows:

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Transwell Chemotaxis,ct leukocyte differentiation (.). Among the proinflammatory activities, chemotaxis in particular has received considerable attention as a target for novel antiinflammatory therapeutics (reviewed in ..).

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Actin Polymerization,to endothelial receptors, increasing adhesion which results in the arrest of the rolling leukocytes, which can then cross the endothelial layer of the blood vessel and enter the tissue using integrins for traction (.–.).

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