Authentication of a Novel Antibody to Zebrafish Collagen Type XI Alpha 1 Chain (Col11a1a)

Jonathon C. Reeck, Makenna J. Hardy, Xinzhu Pu, Cynthia Keller-Peck, Julia Thom Oxford

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Objective: Extracellular matrix proteins play important roles in embryonic development and antibodies that specifically detect these proteins are essential to understanding their function. The zebrafish embryo is a popular model for vertebrate development but suffers from a dearth of authenticated antibody reagents for research. Here, we describe a novel antibody designed to detect the minor fibrillar collagen chain Col11a1a in zebrafish (AB strain). Results: The Col11a1a antibody was raised in rabbit against a peptide comprising a unique sequence within the zebrafish Col11a1a gene product. The antibody was affinity-purified and characterized by ELISA. The antibody is effective for immunoblot and immunohistochemistry applications. Protein bands identified by immunoblot were confirmed by mass spectrometry and sensitivity to collagenase. Col11a1a knockout zebrafish were used to confirm specificity of the antibody. The Col11a1a antibody labeled cartilaginous structures within the developing jaw, consistent with previously characterized Col11a1 antibodies in other species. Col11a1a within formalin-fixed paraffin-embedded zebrafish were recognized by the antibody. The antibodies and the approaches described here will help to address the lack of well-defined antibody reagents in zebrafish research.

Original languageAmerican English
Article number359
JournalBMC Research Notes
Volume14
Issue number1
DOIs
StatePublished - Dec 2021

Keywords

  • Antibody authentication
  • Cartilage
  • Col11a1a
  • Collagen α1(XI)
  • Immunoblot
  • Immunofluorescence microscopy
  • Zebrafish

EGS Disciplines

  • Biochemistry, Biophysics, and Structural Biology
  • Biology

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