PUBLICATION
Authentication of a novel antibody to zebrafish collagen type XI alpha 1 chain (Col11a1a)
- Authors
- Reeck, J.C., Hardy, M.J., Pu, X., Keller-Peck, C., Oxford, J.T.
- ID
- ZDB-PUB-210918-5
- Date
- 2021
- Source
- BMC research notes 14: 359 (Journal)
- Registered Authors
- Hardy, Makenna, Oxford, Julia, Reeck, Jonathon
- Keywords
- Antibody authentication, Cartilage, Col11a1a, Collagen α1(XI), Immunoblot, Immunofluorescence microscopy, Zebrafish
- MeSH Terms
-
- Animals
- Antibodies
- Collagen Type XI*
- Extracellular Matrix Proteins
- Peptides
- Rabbits
- Zebrafish*
- PubMed
- 34526111 Full text @ BMC Res. Notes
Citation
Reeck, J.C., Hardy, M.J., Pu, X., Keller-Peck, C., Oxford, J.T. (2021) Authentication of a novel antibody to zebrafish collagen type XI alpha 1 chain (Col11a1a). BMC research notes. 14:359.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping