{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T01:20:29Z","timestamp":1772587229116,"version":"3.50.1"},"reference-count":52,"publisher":"Portland Press Ltd.","issue":"6","license":[{"start":{"date-parts":[[2019,7,25]],"date-time":"2019-07-25T00:00:00Z","timestamp":1564012800000},"content-version":"vor","delay-in-days":609,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2017,12,22]]},"abstract":"<jats:p>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\u2212\/\u2212 mouse brain and kidney extracts prepared with different detergents, in membrane proteins isolated from the cerebellum, cerebral hemisphere and kidney of WT and Cln3\u2212\/\u2212 mice, in cell extracts of WT and Cln3\u2212\/\u2212 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 different mouse and human proteins, providing a plausible explanation for the lack of specificity of anti-CLN3 antibodies. Our data provide evidence that immunization against a transmembrane protein with low to medium expression level does not necessarily generate specific antibodies. Because of the possible cross-reactivity to other proteins, the specificity of an antibody should always be checked using tissue samples from an appropriate knock-out animal or using knock-out cells.<\/jats:p>","DOI":"10.1042\/bsr20171229","type":"journal-article","created":{"date-parts":[[2017,11,1]],"date-time":"2017-11-01T00:10:49Z","timestamp":1509495049000},"update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Lack of specificity of antibodies raised against CLN3, the lysosomal\/endosomal transmembrane protein mutated in juvenile Batten disease"],"prefix":"10.1042","volume":"37","author":[{"given":"Tarah","family":"Nelson","sequence":"first","affiliation":[{"name":"Pediatric and Rare Diseases Group, Sanford Research, Sioux Falls, South Dakota 57104, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David\u00a0A.","family":"Pearce","sequence":"additional","affiliation":[{"name":"Pediatric and Rare Diseases Group, Sanford Research, Sioux Falls, South Dakota 57104, U.S.A."},{"name":"Department of Pediatrics, Sanford School of Medicine, University of South Dakota Sioux Falls, South Dakota 57104, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Attila\u00a0D.","family":"Kov\u00e1cs","sequence":"additional","affiliation":[{"name":"Pediatric and Rare Diseases Group, Sanford Research, Sioux Falls, South Dakota 57104, U.S.A."},{"name":"Department of Pediatrics, Sanford School of Medicine, University of South Dakota Sioux Falls, South Dakota 57104, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2017,11,23]]},"reference":[{"key":"2021111618585659600_B1","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1016\/0092-8674(95)90274-0","article-title":"Isolation of a novel gene underlying Batten disease, CLN3","volume":"82","author":"Consortium","year":"1995","journal-title":"Cell"},{"key":"2021111618585659600_B2","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1212\/WNL.0b013e31825f0547","article-title":"New nomenclature and classification scheme for the neuronal ceroid lipofuscinoses","volume":"79","author":"Williams","year":"2012","journal-title":"Neurology"},{"key":"2021111618585659600_B3","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1111\/j.1750-3639.2004.tb00499.x","article-title":"Current state of clinical and morphological features in human NCL","volume":"14","author":"Goebel","year":"2004","journal-title":"Brain Pathol."},{"key":"2021111618585659600_B4","doi-asserted-by":"crossref","first-page":"e102593","DOI":"10.1371\/journal.pone.0102593","article-title":"FRET-assisted determination of CLN3 membrane topology","volume":"9","author":"Ratajczak","year":"2014","journal-title":"PLoS One"},{"key":"2021111618585659600_B5","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1006\/mgme.1999.2837","article-title":"Analysis of intracellular distribution and trafficking of the CLN3 protein in fusion with the green fluorescent protein in vitro","volume":"66","author":"Kida","year":"1999","journal-title":"Mol. 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