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Enhanced validation of antibodies for research applications
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science. KTH, Centres, Science for Life Laboratory, SciLifeLab.
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2018 (English)In: Nature Communications, ISSN 2041-1723, E-ISSN 2041-1723, Vol. 9, article id 4130Article in journal (Refereed) Published
Abstract [en]

There is a need for standardized validation methods for antibody specificity and selectivity. Recently, five alternative validation pillars were proposed to explore the specificity of research antibodies using methods with no need for prior knowledge about the protein target. Here, we show that these principles can be used in a streamlined manner for enhanced validation of research antibodies in Western blot applications. More than 6,000 antibodies were validated with at least one of these strategies involving orthogonal methods, genetic knockdown, recombinant expression, independent antibodies, and capture mass spectrometry analysis. The results show a path forward for efforts to validate antibodies in an application-specific manner suitable for both providers and users.

Place, publisher, year, edition, pages
Nature Publishing Group, 2018. Vol. 9, article id 4130
National Category
Immunology in the medical area
Identifiers
URN: urn:nbn:se:kth:diva-237096DOI: 10.1038/s41467-018-06642-yISI: 000446566000016PubMedID: 30297845Scopus ID: 2-s2.0-85054574300OAI: oai:DiVA.org:kth-237096DiVA, id: diva2:1259688
Funder
Science for Life Laboratory - a national resource center for high-throughput molecular bioscienceKnut and Alice Wallenberg Foundation
Note

QC 20181030

Available from: 2018-10-30 Created: 2018-10-30 Last updated: 2018-10-30Bibliographically approved

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Edfors, FredrikMaddalo, GianlucaSivertsson, ÅsaTegel, HannaHober, SophiaAl-Khalili Szigyarto, CristinaFagerberg, Linnvon Feilitzen, KalleUhlén, Mathias

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Edfors, FredrikHober, AndreasLinderbäck, KlasMaddalo, GianlucaSivertsson, ÅsaTegel, HannaHober, SophiaAl-Khalili Szigyarto, CristinaFagerberg, Linnvon Feilitzen, KalleOksvold, PerForsström, BjörnUhlén, Mathias
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Protein ScienceScience for Life Laboratory, SciLifeLab
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Immunology in the medical area

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