By Margaret A. Scofield, Jean D. Deupree, David B. Bylund (auth.), Curtis A. Machida (eds.)
In Adrenegic Receptor Protocols, Curtis Machida and a panel of specialist investigators current a complete number of sleek molecular equipment for studying adrenergic receptors and corresponding moment messenger platforms. those confirmed and easily reproducible ideas make the most of genetic, RNA, protein expression, transactivator, and moment messenger methodologies, in addition to immunocytochemical, electrophysiological, transgenic, and in situ hybridization ways. all the specialists writing right here info using their selected strategy in studying the adrenergic receptor procedure, utilizing features of the genetic circulation of data as a advisor: DNA ' RNA ' transactivator ' protein expression ' moment messenger analyses ' mobile analyses ' transgenic complete animal approaches.
complete and wealthy in useful aspect, Adrenergic Receptor Protocols offers the 1st selection of reproducible tools for the learn of those very important regulators of CNS-mediated habit and neural functionality. Its cutting-edge tools represent modern-day optimum reference for all neurobiologists, neurochemists, neurologists, and pharmacologists learning this incredibly vital classification of receptors.
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17. , Kingston, R. , Moore, D. , Seidman, J. , Smith, J. , et al. (1992) Production of a complete cDNA library, in Current Protocols in Molecular Biology, vol. 1. Wiley Interscience, New York, pp. 8. 18. Frohman, M. A. (1990) RACE: rapid amplificiation of cDNA ends, in PCR Protocols—A Guide to Methods and Applications (Innis, M. , Gelfand, D. , Sninsky, J. , and White, T. ), Academic, San Diego, CA, pp. 28–38. Sequence Analysis 53 3 Analyses of Adrenergic Receptor Sequences Jean D. Deupree, Margaret A.
11. Dilute this positive aliquot to 300 colonies/mL by increasing the volume 100fold with the addition of approx 10 mL with LB/kanamycin media. Receptor Gene Isolation 41 12. Aliquot 100 µL (30 colonies) of the diluted mixture into additional 96-well microtiter dishes. 13. Repeat steps 7–9 above to identify a positive clone. 14. Plate the entire volume of the well with the positively identified clone on LB agar/kanamycin plates. 15. Toothpick each colony into individual wells of the 96-well microtiter dishes containing 100 µL of LB/kanamycin media, and grow for 6–8 h.
Minneman, K. , et al. (1994) IV. International Union of Pharmacology Nomenclature of Adrenoceptors. Pharmacol. Rev. 46, 121–136. 6. Johnson, R. D. and Minneman, K. P. (1987) Differentiation of α1-adrenergic receptors linked to phosphatidylinositol turnover and cyclic AMP accumulation in rat brain. Mol. Pharmacol. 31, 239–246. 7. Minneman, K. , and Abel, P. W. (1988) Comparison of α1-adrenergic receptor subtypes distinguished by chloroethylclonidine and WB4101. Mol. Pharmacol. 33, 509–514. 8. Morrow, A.
Adrenergic Receptor Protocols by Margaret A. Scofield, Jean D. Deupree, David B. Bylund (auth.), Curtis A. Machida (eds.)