By Proudfoot, Amanda E.I.; Wells, Timothy N.C.; Power, Christine
Professional investigators describe intimately the main commonplace concepts in chemokine biology. protecting either ligands and receptors, those without difficulty reproducible equipment disguise all features of chemokine study, starting from the cloning and characterization of chemokines and their receptors, by using animal versions to check chemokine functionality in vivo. each one strategy additionally comprises appropriate historical past details, in addition to supplying an invaluable bibliography that renders the examine of chemokines available in any respect degrees of expertise. accomplished and hugely functional, Chemokine Protocols bargains experimental and scientific chemokine researchers trendy gold-standard number of confirmed tools for examining this biologically ubiquitous and significant type of proteins.
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Extra info for Chemokine Protocols (Methods in Molecular Biology Vol 138)
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1990) pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 18, 5322. 10. Maruyama, K. and Sugano, S. (1994) Oligo-capping: a simple method to replace the cap structure of eukaryotic mRNAs with oligoribonucleotides. Gene 138, 171–174. 11. , and Yoshie, O. (1995) Molecular cloning of a novel C or gamma type chemokine, SCM-1. FEBS Lett. 360, 155–159. 12. Klein, R. , and Rosenthal, A. (1996) Selection for genes encoding secreted proteins and receptors. Proc. Natl. Acad. Sci. USA 93, 7108–7113.
High-Five cells are relatively resistant for pipeting and easily detached with pipeting when cells are nearly confluent. But in subconfluent, both cells are strongly attached to the flask and detachment of these cells may reduce the viability. 5. Usually, we do not determine the titer of recombinant virus, since this protocol works well for over 10 chemokines. Successful infection is easily confirmed by the morphology of the High Five cells. Uninfected cells are firmly attached and infected cells become round up.
177, 1497–1504. 9. Lopez, P. , and Dreyfus, M. (1994) The use of a tRNA as a transcriptional reporter: the T7 late promoter is extremely efficient in Escherichia coli but its transcripts are poorly expressed. Nucleic Acids Res. 22, 1186–1193. 10. Ramesh, V, Amitabha De, and Nagaraja, V. (1994) Engineering hyperexpression of bacteriophage Mu protein C by removal of secondary structure at the translation initiation region. Prot. Engineer. 7, 1053–1057. 11. Kane, J. F. (1995) Effects of rare codon clusters on high-level expression of heterologous protein in Escherichia coli.