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Synthesis of biotin-labelled dexamethasone derivatives. Novel hormone-affinity probes.

Authors
  • Manz, B
  • Heubner, A
  • Köhler, I
  • Grill, H J
  • Pollow, K
Type
Published Article
Journal
European journal of biochemistry / FEBS
Publication Date
Mar 15, 1983
Volume
131
Issue
2
Pages
333–338
Identifiers
PMID: 6832154
Source
Medline
License
Unknown

Abstract

A new, general methodology for 'sandwich' affinity chromatography of steroid hormone receptors is proposed, the part purification of the human spleen tumor glucocorticoid receptor is quoted as an illustration. 9-Fluoro-16 alpha-methyl-11 beta, 17-dihydroxy-1,4-androstadiene-3-one-17 beta-carboxylic acid was coupled to biotin using pentamethylenediamine (BioDex 1) as a spacer. The bifunctional derivative binds to glucocorticoid receptors and avidin-Sepharose and efficiently protects the glucocorticoid receptor against inactivation when previously added during homogenisation. We have standardized the capacity and optimum conditions for elution of receptor-BioDex-1 complexes which are bound to avidin-Sepharose. Receptor purification of several thousand fold can be obtained with good yield.

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