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Separation of alpha and beta chains of rabbit globin using SDS-polyacrylamide gel electrophoresis

✍ Scribed by T.Gordon Wood; Joseph R. Shaeffer

Publisher: Elsevier Science
Year: 1975
Tongue: English
Weight: 361 KB
Volume: 63
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(75)90197-9

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✦ Synopsis

A procedure to separate the (Y and /3 globin chains of rabbit hemoglobin. denatured with sodium dodecyl sulfate in the presence of mercaptoethanol. on a column of polyacrylamide gel was developed. The identity of the two separated chains was verified by (a) differences in distribution of radioactivity between the chains when the hemoglobin samples were labeled uniformly with various aH-or 14C-labeled amino acids; (b) the analysis of the chain distribution of radioactivity in purified hemoglobin isolated from rabbit reticulocytes. pulse-labeled with [3H] leucine: and (c) the separation pattern of a mixture of authentic [~x-~H]~ and [p-"C]-labeled globin chains. The globin chains of human hemoglobin A also could be separated in a similar manner. This procedure is particularly useful when only mrcrogram quantities of hemoglobin are available for study. 135

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