Download Chemokines by Richard Horuk (Eds.) PDF
By Richard Horuk (Eds.)
Chemokines are significant mediators of immune cells and are occupied with quite a lot of proinflammatory human illnesses, together with rheumatoid arthritis, a number of sclerosis, and organ transplant rejection. It has lately been came across that their receptors are desirous about HIV an infection. The characterization of those molecules and their receptors is hence of basic significance in figuring out a few human ailments and infections. This quantity and its spouse Volume 288 on Chemokine Receptors provide complete experimental protocols utilized in this box of study.
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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.
Twenty microliters of a reduction/alkylation mixture [2 /zl of tri-n-butylphosphine, 5 /~1 of 4-vinylpyridine (Aldrich, Milwaukee, WI), and 100/~1 of acetonitrile] is added to the sample, and the cartridge is immediately reassembled. The reaction cartridge is equilibrated with N-methylpiperidine, and excess reagents are removed by washing with n-butyl chloride and ethyl acetate. This reaction cycle is followed by the normal reaction cycles. Cysteine residues can also be alkylated in solution with acrylamide.
Dixon, Anal. Biochern. 161, 524 (1987). ~xD. C. Brune, Anal. Biochem. 207, 285 (1992). 5 78% 16% 6% AATELRXVXLTVTPK 15% ATELRXVXLTVTP 25% TELRXVXLTVTPKINPKLIANXEVIPXGXQ ELRXVXLXXTPKXNXXXXA 44% 16~ Fl~. 3. Identification of neutrophil chemotactic proteins as GCP-2 isoforms by NH~terminal amino acid sequence analysis. (A) Four 6-kDa proteins purified by R P - H P L C from the conditioned m e d i u m of h u m a n osteosarcoma cells and corresponding to neutrophil chemotactic activity (fractions 54~ 56, 60, and 67, see Fig.