Validation of anti-glucocerebrosidase antibodies for western blot analysis on protein lysates of murine and human cells.

Abstract

Gaucher disease (GD) is a rare lysosomal storage disorder caused by mutations in the GBA1 gene, encoding the lysosome-resident glucocerebrosidase enzyme involved in the hydrolysis of glucosylceramide. The discovery of an association between mutations in GBA1 and the development of synucleinopathies, including Parkinson disease, has directed attention to glucocerebrosidase as a potential therapeutic target for different synucleinopathies. These findings initiated an exponential growth in research and publications regarding the glucocerebrosidase enzyme. The use of various commercial and custom-made glucocerebrosidase antibodies has been reported, but standardized in-depth validation is still not available for many of these antibodies. This work details the evaluation of several previously reported glucocerebrosidase antibodies for western blot analysis, tested on protein lysates of murine gba+/+ and gba-/- immortalized neurons and primary human wild-type and type 2 GD fibroblasts.

• Authors: Wenduo Qi, Brad A Davidson, Matthew Nguyen, Taylor Lindstrom, Richard J Grey, Robert Burnett, Elma Aflaki, Ellen Sidransky, Wendy Westbroek
• Journal: The Biochemical journal (Biochem J) Vol. 476 Issue 2 Pg. 261-274 (01 25 2019) ISSN: 1470-8728 [Electronic] England
• PMID: 30578288 (Publication Type: Journal Article, Research Support, N.I.H., Intramural, Validation Study)
• Copyright: © 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Chemical References

• Antibodies
• GBA protein, human
• Glucosylceramidase

Topics

• Animals
• Antibodies (chemistry)
• Blotting, Western
• Cell Line, Transformed
• Fibroblasts (enzymology, pathology)
• Gaucher Disease (enzymology, genetics, pathology)
• Glucosylceramidase (genetics, metabolism)
• Humans
• Mice
• Mice, Knockout
• Parkinson Disease (enzymology, genetics, pathology)