By Richard Horuk (Eds.)
Chemokines are significant mediators of immune cells and are excited by quite a lot of proinflammatory human illnesses, together with rheumatoid arthritis, a number of sclerosis, and organ transplant rejection. It has lately been found that their receptors are focused on HIV an infection. The characterization of those molecules and their receptors is hence of basic significance in knowing a couple of human ailments and infections. This quantity and its significant other Volume 288 on Chemokine Receptors provide accomplished experimental protocols utilized in this box of study.
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G. Garnham, D. A. Bergman, and P. F. Alewood, Tetrahedron Lett. 36, 8871 (1995). m M. Baca, T. W. Muir, M. Schn61zer, and S. B. H. Kent, J. A m . Chem. Soc. 117, 1881 (1995). u L. E. Canne, A. R. Ferre-D'Amare, S. K. Burley, and S. B. H. Kent, J. A m . Chem. Soc. 117, 2998 (1992). 12p. E. Dawson, T. W. Muir, I. Clark-Lewis, and S. B. H. Kent, Science 266, 776 (1994). METHODS IN ENZYMOLOGY, VOL. 287 Copyright © 1997 by Academic Press All rights of reproduction in any form reserved. 3 II C-N H'CH-c - ~ _ , , ~ " ~ - .
Willman, B. Griffith, D. D. Von Hoff, and 1. B. Roninson, Proc. Natl. Acad. Sci. A. 87, 7160 (1990). 23 L. D. Murphy, C. E. Herzog, J. B. Rudick, A. T. Fojo, and S. E. Bates, Biochemistry 29, 10351 (1990).  GRANULOCYTE CHEMOTACTIC PROTEIN-2 1SOFORMS 33 has been demonstrated in our laboratory for the quantitation of other cytokine m R N A s . 24 In the latter study, the quantitation was even p e r f o r m e d with an internal control to obtain absolute values of m R N A molecules. The method described here is especially useful for the quantitation of cytokine m R N A molecules because these are present at low abundance and have a short half-life.
D. Baly, U. Gibson, D. Allison. and L. DeForge, Methods EnzymoL 287, , 1997 (this volume). ~1S. R. McColl and P. H. ~ Enzymol. 288,  (1997). ~ ~'Counts represent the average of six replicate wells per dose. The addition of 10% serum resuited in the incorporation of 33,440 _+ 1,639 cpm of [3H]thymidine into the cells. [SH]Thymidine Incorporation Assay for Mitogenesis T h e [3H]thymidine p r o l i f e r a t i o n assay, which m e a s u r e s [3H]thymidine i n c o r p o r a t i o n into D N A , is a s i m p l e assay to m e a s u r e t h e b i o l o g i c a l activity of M G S A .