Validation of Anti-Glucocerebrosidase Antibodies for Western Blot Analysis on Protein Lysates of Murine and Human Cells

Research Article | Biochemical Journal
Validation of Anti-Glucocerebrosidase Antibodies for Western Blot Analysis on Protein Lysates of Murine and Human Cells
Author's Information

Wenduo Qi

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Brad A. Davidson

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Matthew Nguyen

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Taylor Lindstrom

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Richard J. Grey

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Robert Burnett

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Elma Aflaki

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Ellen Sidransky

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.

Wendy Westbroek

Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, U.S.A.
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.

Keywords

antibody validation, Gaucher disease, glucocerebrosidase, knockout mice, Parkinson's disease

DOI: 10.1042/BCJ20180708

ADLID: 95454-v4

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Keywords
antibody validation Gaucher disease glucocerebrosidase knockout mice Parkinson's disease

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