Chemokine Protocols

1. Front Matter

   Pages i-xiii

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2. Purification of Chemokines from Natural Sources

   • Jens-M. Schröder

   Pages 1-10

3. Cloning of Novel Chemokines Using a Signal Sequence Trap Method

   • Toshio Imai

   Pages 11-21

4. Chemokine Expression in Insect Cells

   • Toshio Imai

   Pages 23-32

5. Expression of Chemokines in Escherichia coli

   • Michael D. Edgerton, Lars-Ole Gerlach, Thomas P. Boesen, Bernard Allet

   Pages 33-40

6. Expression of Chemokines in the Periplasmic Space of E. coli

   • Jochen Pfirstinger, Matthias Mack

   Pages 41-45

7. Synthesis of Chemokines

   • Ian Clark-Lewis

   Pages 47-63

8. Identification of Novel Chemokines From Expressed Sequence Tag Databases

   • Timothy N. C. Wells, Manuel C. Peitsch

   Pages 65-73

9. Purification of Recombinant Chemokines from E. coli

   • Amanda E. I. Proudfoot, Frédéric Borlat

   Pages 75-87

10. Chemokine Receptor Cloning

    • Philip M. Murphy

    Pages 89-98

11. Generation of Stable Cell Lines Expressing Chemokine Receptors

    • Christine A. Power, Alexandra Meyer

    Pages 99-104

12. Modified Microchemotaxis Assays

    • Dennis D. Taub

    Pages 105-112

13. Transwell Chemotaxis

    • Paul D. Ponath, Juan Wang, Heidi Heath

    Pages 113-120

14. Endothelial Cell Chemotaxis Assays

    • Darcey Black

    Pages 121-127

15. Radiolabeled Chemokine Binding Assays

    • Bruce L. Daugherty, Salvatore J. Siciliano, Martin S. Springer

    Pages 129-134

16. Scintillation Proximity Binding Assay

    • Sami Alouani

    Pages 135-141

17. Calcium Mobilization

    • Raphaële Buser, Amanda E. I. Proudfoot

    Pages 143-148

18. Actin Polymerization

    • Jörn Elsner, Alexander Kapp

    Pages 149-152

19. Reactive Oxygen Release

    • Jörn Elsner, Alexander Kapp

    Pages 153-156

20. Histamine Release

    • Michael A. Lett-Brown, Rafeul Alam

    Pages 157-162

The chemokines family of small proteins are involved in numerous b- logical processes ranging from hematopoiesis, angiogenesis, and basal l- kocyte trafficking to the extravasation and tissue infiltration of leukocytes in response to inflammatory agents, tissue damage, and bacterial or viral infection. Chemokines exert their effects through a family of seven G-protein coupled transmembrane receptors. Worldwide interest in the chemokine field surged dramatically early in 1996, with the finding that certain chemokine receptors were the elusive coreceptors, required along with CD4, for HIV infection. Today, though over 40 human chemokines have been described, the n- ber of chemokine receptors lags behind—only 17 human chemokine receptors have been identified so far. What has emerged over the years is that most chemokine receptors bind several distinct ligands, and indeed the majority of chemokines are able to bind to multiple chemokine receptors, explaining to some extent the apparent disparity in the numbers of chemokines and rec- tors. Yet in spite of the apparent redundancy in chemokine/chemokine rec- tor interactions, it is clear that in vivo, spatial, temporal, and indeed cell- and tissue-specific expression of both chemokines and their receptors are imp- tant factors in determining the precise nature of cellular infiltrates in phy- ological and pathological processes.

highly recommended...Seldom does on find so much valuable information, concisely presented and with such obvious potential for the future, made available in one volume." - E-Streams

"...Chemokine Protocols is a collection of clear, well written protocols and I would recommend it as an excellent starting point for any scientist with an interest in chemokines." - Molecular Biotechnology

• Serono Pharmaceutical Research Institute, Geneva, Switzerland

  Amanda E. I. Proudfoot, Timothy N. C. Wells, Christine A. Power

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