Authentication of a novel antibody to zebrafish collagen type XI alpha 1 chain (Col11a1a)
Autor: | Makenna J. Hardy, Jonathon C. Reeck, Cynthia Keller-Peck, Julia Thom Oxford, Xinzhu Pu |
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Rok vydání: | 2021 |
Předmět: |
Science (General)
QH301-705.5 Collagen Type XI Immunoblot Immunofluorescence microscopy Antibodies General Biochemistry Genetics and Molecular Biology Extracellular matrix Gene product Q1-390 03 medical and health sciences 0302 clinical medicine medicine Animals Collagen α1(XI) Biology (General) Zebrafish 030304 developmental biology Extracellular Matrix Proteins 0303 health sciences biology Chemistry General Medicine biology.organism_classification Collagen type XI alpha 1 3. Good health Cell biology Research Note Cartilage Col11a1a Antibody authentication biology.protein Collagenase Medicine Immunohistochemistry Rabbits Antibody Peptides 030217 neurology & neurosurgery Function (biology) medicine.drug |
Zdroj: | BMC Research Notes BMC Research Notes, Vol 14, Iss 1, Pp 1-8 (2021) |
ISSN: | 1756-0500 |
Popis: | 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. |
Databáze: | OpenAIRE |
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