Generation of monoclonal antibody against canine neural-cell adhesion molecule

Area of Science:

• Immunology
• Neuroscience
• Molecular Biology

Background:

• Neural-cell adhesion molecule (N-CAM) plays a crucial role in neuronal development and function.
• Understanding canine N-CAM expression and function is vital for comparative neuroscience and translational research.

Purpose of the Study:

• To generate and characterize a novel monoclonal antibody, K9BYU, specific for canine neural-cell adhesion molecule (N-CAM).
• To evaluate the utility of K9BYU as a research tool for studying canine N-CAM.

Main Methods:

• Generation of monoclonal antibody K9BYU using recombinant canine N-CAM extracellular domain expressed in Escherichia coli.
• Immunoreactivity confirmation via Western blotting against insect cell-expressed canine N-CAM.
• Analysis of canine brain tissue using Western blotting to detect N-CAM isoforms.

Main Results:

• K9BYU demonstrated immunoreactivity to insect cell-expressed canine N-CAM.
• Western blotting of canine brain tissue revealed that K9BYU detects three major N-CAM isoforms (N-CAM-120, -140, and -180).
• These detected isoforms correspond to known major isoforms in human and mouse N-CAM.

Conclusions:

• The monoclonal antibody K9BYU is a specific and effective tool for detecting canine N-CAM.
• K9BYU facilitates the study of canine N-CAM isoforms, aiding comparative neuroscience research.