Lack of specificity of antibodies raised against CLN3, the lysosomal/endosomal transmembrane protein mutated in juvenile Batten disease

Bioscience Reports
Tarah NelsonAttila D Kovács

Abstract

Juvenile CLN3 (Batten) disease, a fatal, childhood neurodegenerative disorder, results from mutations in the CLN3 gene encoding a lysosomal/endosomal transmembrane protein. The exact physiological function of CLN3 is still unknown and it is unclear how CLN3 mutations lead to selective neurodegeneration. To study the tissue expression and subcellular localization of the CLN3 protein, a number of anti-CLN3 antibodies have been generated using either the whole CLN3 protein or short peptides from CLN3 for immunization. The specificity of these antibodies, however, has never been tested properly. Using immunoblot experiments, we show that commercially available or researcher-generated anti-CLN3 antibodies lack specificity: they detect the same protein bands in wild-type (WT) and Cln3-/- mouse brain and kidney extracts prepared with different detergents, in membrane proteins isolated from the cerebellum, cerebral hemisphere and kidney of WT and Cln3-/- mice, in cell extracts of WT and Cln3-/- mouse embryonic fibroblast cultures, and in lysates of BHK cells lacking or overexpressing human CLN3. Protein BLAST searches with sequences from peptides used to generate anti-CLN3 antibodies identified short motifs present in a number of diffe...Continue Reading

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Citations

Mar 11, 2020·JIMD Reports·Willemijn F E KuperPeter M van Hasselt
Oct 20, 2020·Acta Ophthalmologica·Willemijn F E KuperMaria M van Genderen
Oct 28, 2019·Biochimica Et Biophysica Acta. Molecular Basis of Disease·Christopher J MinnisTristan R McKay

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Methods Mentioned

BETA
glycosylation
protein assay
Protein

Software Mentioned

Protein BLAST
Blast

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