By P. Michael Conn (Eds.)
This quantity plus its spouse, quantity 294, provides leading edge strategies for the research of ion channels, starting from the molecular to the physiological point. those volumes complement quantity 207 of Methods inEnzymology
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Extra resources for Ion Channels Part B, 1st Edition
M o r e o v e r , one can engineer a particular function of a channel to reveal its prior history, namely, to learn something about the assembly of individual subunits, In this regard the strategy and m e t h o d s described later can be used to answer the following questions: D o e s synthesis and assembly of different channel subunits occur in the same shared compartment'? Are > Z. G. Galen and R. E. Oslwald. ,1. Biol. Chem. 270, 20(/0 (1995). _'t M. Holman, C. Maron, and S. Heinemann, Neuron 13, 1331 (1994).
Determine the doubling rate at log phase and the O D at stationary stage. ) |0 ASSEMBLY [11 Testing Domain Interactions by t3-Gal Assay Two protocols have been commonly used to test/3-Gal activity. Here we describe the procedure for performing the/3-Gal assay on a membrane support. Refer to Current Protocols for the liquid culture/3-Gal assay. 9 Screen Yeast Colonies with/3-Gal Assay on a Nitrocelhdose Filter l. Wear gloves for all steps. Label the nitrocellulose filter with a fine marker. Carefully place the filter onto the plate to be assayed.
Sheng. W. Skach, D. Pain, and C. Dcutsch, ,I. Biol. Chem. 271, 18904 (1996). ~Z. Shcng, W. Skach. V. Santarelli, and C. Deutsch, Biochem. 36, 15501 15513 (1997). YOF ION C H A N N E L S 19 wheat germ agglutinin (WGA) systems, which contain cellular components necessary for protein synthesis (tRNA, ribosomes, amino acids, and initiation, elongation, and termination factors) and are capable of a variety of posttranslational processing activities (acetylation, isoprenylation, proteolysis, and some phosphorylation activity).