A simple method for the preparation and purification of C1 complement cleaved beta 2-microglobulin from human serum.
J Immunol Methods
; 205(1): 29-33, 1997 Jun 23.
Article
em En
| MEDLINE
| ID: mdl-9236912
ABSTRACT
A simple method is described for the preparation of proteolytically processed forms of beta 2-microglobulin suitable for structural and biological studies. PEG 6000 was added to the serum of healthy individuals to precipitate the C1 complement complex from C1 esterase inhibitor (C1-inh). After dissolving the precipitate containing the C1 complement in Tris-HCl buffer, pH 7.6, efficient conversion of added beta 2-microglobulin to desLys58 beta 2-microglobulin was observed. Addition of a specific carboxypeptidase B inhibitor (Plummers inhibitor) could partly prevent the deletion of Lys-58 from cleaved beta 2-microglobulin, whereby Lys58-cleaved beta 2-microglobulin was obtained. The proteolytically processed forms were subsequently purified by G-75 Sephadex gel filtration followed by chromatofocusing. A yield of 10-40% of proteolytically processed beta 2-microglobulin was obtained. Only one component was seen by SDS-PAGE stained with Coomassie Brilliant Blue.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Complemento C1
/
Microglobulina beta-2
Limite:
Humans
Idioma:
En
Revista:
J Immunol Methods
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Dinamarca