U.S. flag

An official website of the United States government

Dot gov

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Https

Secure .gov websites use HTTPS
A lock ( ) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.

PubAg

Main content area

Development of rabbit monoclonal antibodies for detection of alpha-dystroglycan in normal and dystrophic tissue

Author:
Marisa J. Fortunato, Dan J. Nonneman, Charlotte E. Ball, Katrin Hollinger, Niraj B. Patel, Jill N. Modi, Vedika Rajasekaran, Jason W. Ross, Eileen J. Kennedy, Joshua T. Selsby, Aaron M. Beedle
Source:
Plos One 2014 v.9 no.5 pp. e97567
Subject:
Western blotting, animal models, cytoskeletal proteins, epitopes, extracellular matrix, fluorescent antibody technique, glycosylation, laminin, metastasis, mice, molecular weight, monoclonal antibodies, muscles, muscular dystrophy, neoplasms, rabbits, rats, skeletal muscle, swine
Abstract:
Alpha-dystroglycan requires a rare O-mannose glycan modification to form its binding epitope for extracellular matrix proteins such as laminin. This functional glycan is disrupted in a cohort of muscular dystrophies, the secondary dystroglycanopathies, and is abnormal in some metastatic cancers. The most commonly used reagent for detection of alpha-dystroglycan is mouse monoclonal antibody IIH6, but it requires the functional O-mannose structure for recognition. Therefore, the ability to detect alpha-dystroglycan protein in disease states where it lacks the full O-mannose glycan has been limited. To overcome this hurdle, rabbit monoclonal antibodies against the alpha-dystroglycan C-terminus were generated. The new antibodies, named 5–2, 29–5, and 45–3, detect alpha-dystroglycan from mouse, rat and pig skeletal muscle by Western blot and immunofluorescence. In a mouse model of fukutin-deficient dystroglycanopathy, all antibodies detected low molecular weight alpha-dystroglycan in disease samples demonstrating a loss of functional glycosylation. Alternately, in a porcine model of Becker muscular dystrophy, relative abundance of alpha-dystroglycan was decreased, consistent with a reduction in expression of the dystrophin-glycoprotein complex in affected muscle. Therefore, these new rabbit monoclonal antibodies are suitable reagents for alpha-dystroglycan core protein detection and will enhance dystroglycan-related studies.
Agid:
59658
Handle:
10113/59658