Isolation of human type X collagen and immunolocalization in fetal human cartilage

Thorsten KIRSCH, Klaus VON DER MARK

Research output: Contribution to journalArticlepeer-review

Abstract

Type X Collagen was extracted with 1 M NaCl and 10 mM dithiothreitol at neutral pH from fetal human growth plate cartilage and purified to homogeneity by gel filtration and anion‐exchange chromatography. The purified protein migrates in SDS/polyacrylamide gels with an apparent Mr of 66000 under reducing conditions, and as a high‐Mr oligomer under non‐reducing conditions. Purified collagenase digests most of the molecule; pepsin digestion at 4°C decreases the Mr of the monomer to 53000. A rabbit antiserum was raised against purified human type X collagen; the IgG fraction was specific for this collagen by criteria of ELISA and immunoblotting after absorption with collagen types I, II, VI, IX and XI. Immunohistological studies localized type X collagen exclusively in the zone of hypertrophic and calcifying cartilage.

Original languageEnglish (US)
Pages (from-to)575-580
Number of pages6
JournalEuropean Journal of Biochemistry
Volume196
Issue number3
DOIs
StatePublished - Mar 1991

ASJC Scopus subject areas

  • Biochemistry

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