By John N. Abelson, Melvin I. Simon, David Barnes, Jennie P. Mather, Gordon H. Sato
This quantity contains sections facing particular development components during which new, up-to-date, or substitute systems are provided for purification, bioassay, radiolabeling and radioreceptor assay, immunoassay, receptor id, and quantification and comprises common suggestions for the learn of development components. For these surprising with a specific sector resembling the oncogene-growth issue courting orientation chapters were incorporated
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Extra resources for Peptide Growth Factors Part C
Sample text
After 5-7 days at room temperature, the slides are developed in D-19 developer at 20° for 4 min, fixed in rapid fix for 2 min, and washed in running water at least 20 min before being air dried. The developed film is analyzed using computerized densitometry. To identify the brain or embryo structures, reference atlases 8,9 are used. To identify neuroanatomical sites or specific organs, sections can be stained with routine histological staining techniques, for example, thionin for brain tissue and hematoxylin and eosin for chick embryo tissues.
The eluted material from the gel-filtration step consists of native, monomeric growth factor and some contaminants, consisting of modified forms of the growth factor itself. Several such modifications have been identified and characterized, 36'43'44such as variants in which the internal methionine residue has been oxidized to a methionine sulfoxide (IGF-I, tlGF-I), variants in which misincorporation of norleucine for methionine has occurred during fermentation (IGF-I, tlGF-I), forms with incorrectly paired disulfide bonds, and proteolytically degraded forms (IGF-I, tlGF-I, and IGF-II).
Neurosci. 2, 1129 (1982). 5 M. Herkenham, in "Brain Receptor Methodologies" (P. J. Marangos, I. C. Campbell, and R. M. ), p. 127. Academic Press, New York, 1984. 6 M. Herkenham, in "Molecular Neuroanatomy" (S. W. Van Leeuwen, R. M. Buijs, C. W. Pool, and O. ), p. 111. Elsevier, Amsterdam, 1988. METHODS IN ENZYMOLOGY, VOL. 198 26 IGF, NGF, AND PDGF [3] (Gly-976 to Gly-981 and Lys-1003). The entire unmodified IGF-I proreceptor polypeptide chain having a predicted M r of 151,869 can thus be subdivided into the 80,423 a subunit that lacks sequences with membranespanning characteristics, and a 70,866/3 subunit that contains tyrosine kinase-homologous sequences and appears to anchor the intact receptor complex in the plasma membrane.