A recombination assay of high molecular weight nerve growth factor protein complexes

H. D. Shine, J. R. Perez Polo

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The 7S Nerve Growth Factor (NGF) found in mouse is a protein composed of three dissimilar subunits designated α, γ, and β. Mouse NGF reversibly dissociates into its component subunits at pH values lower than 5 or greater than 8. The 7S protein complex is in fast equilibrium with the free subunits. A quantitative determination of 7S NGF based on the reversible dissociation of the molecule into its subunits is reported here. The basis for this procedure is the addition of [125I]α subunit which competes with the native alpha in the 7S complex. The level of NGF present in a sample can be measured in terms of the displacement of [125I]α from the α to the NGF position in a linear sucrose gradient. Measurements are sensitive to 3.5 ng of NGF, linear, and specific for the NGF molecule. This exchange is unaffected by competing unrelated growth factors and inhibiting enzymes. Its specificity was checked by the standard bioassay and by radoimmunoassay. In addition, the assay technique overrides masking by biological inhibitors and/or by the presence of antibodies directed against the IgG contaminant in murine 7S NGF.

Original languageEnglish (US)
Pages (from-to)513-519
Number of pages7
JournalJournal of Neurochemistry
Volume26
Issue number3
StatePublished - 1976
Externally publishedYes

Fingerprint

Nerve Growth Factor
Genetic Recombination
Assays
Molecular Weight
Molecular weight
Proteins
Molecules
Bioassay
Biological Assay
Sucrose
Intercellular Signaling Peptides and Proteins
Immunoglobulin G
Impurities
Antibodies
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

A recombination assay of high molecular weight nerve growth factor protein complexes. / Shine, H. D.; Perez Polo, J. R.

In: Journal of Neurochemistry, Vol. 26, No. 3, 1976, p. 513-519.

Research output: Contribution to journalArticle

Shine, H. D. ; Perez Polo, J. R. / A recombination assay of high molecular weight nerve growth factor protein complexes. In: Journal of Neurochemistry. 1976 ; Vol. 26, No. 3. pp. 513-519.
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