Preparation and characterization of an immuno-electron microscope tracer consisting of a heme-octapeptide coupled to fab.

Détails

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Accès restreint UNIL
Etat: Public
Version: Final published version
Licence: Non spécifiée
ID Serval
serval:BIB_7C1D0DB5CCCD
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Preparation and characterization of an immuno-electron microscope tracer consisting of a heme-octapeptide coupled to fab.
Périodique
The Journal of experimental medicine
Auteur⸱e⸱s
Kraehenbuhl J.P., Galardy R.E., Jamieson J.D.
ISSN
0022-1007 (Print)
ISSN-L
0022-1007
Statut éditorial
Publié
Date de publication
01/01/1974
Peer-reviewed
Oui
Volume
139
Numéro
1
Pages
208-223
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
A heme-octapeptide (mol wt 1,550) has been obtained from cytochrome c by successive pepsin and trypsin hydrolysis and purified by gel filtration and countercurrent distribution. It possesses peroxidatic activity characterized by an apparent K(m) of 0.2 M, an apparent v(max) of 4 mmol/min per mg of peptide, and a pH optimum of 7.0. Using a novel two-step conjugation procedure, the heme-octapeptide was coupled to rabbit Fab antibody fragments by first derivatizing it with the N-hydroxysuccinimide ester of p-formylbenzoic acid and subsequently allowing it to form a Schiff base with the amino groups of Fab. Stable covalent linkages were then obtained by reduction of the Schiff bases with sodium borohydride. The conjugate consists of approximately 2 heme-octapeptides attached to each Fab molecule. The molecular weight is 45,000 daltons when coupled to sheep Fab and 50,000 daltons with a Stokes radius of 32 A, when conjugated to rabbit Fab. Its peroxidatic activity is characterized by an apparent K(m) of 0.4 M, an apparent v(max) of 0.4 mmol/min and per mg of attached heme-octapeptide and a pH optimum of 7.0. The conjugate has been used for the localization at the electron microscope level of secretory immunoglobulins in the mammary gland of lactating rabbits.
Mots-clé
Animals, Cytochrome c Group/analysis, Heme, Horses, Immunochemistry, Immunoglobulin Fab Fragments, Microscopy, Electron, Oligopeptides, Peptides/isolation & purification
Pubmed
Web of science
Open Access
Oui
Création de la notice
25/01/2008 16:06
Dernière modification de la notice
09/08/2024 13:38
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