Enhanced validation of antibodies for research applications

被引：71

作者：

Edfors, Fredrik ^{[1
,2
]}

Hober, Andreas ^{[1
,2
]}

Linderback, Klas ^{[2
]}

Maddalo, Gianluca ^{[1
,2
]}

Azimi, Alireza ^{[3
]}

Sivertsson, Asa ^{[1
,2
]}

Tegel, Hanna ^{[2
]}

Hober, Sophia ^{[2
]}

Szigyarto, Cristina Al-Khalili ^{[1
,2
]}

Fagerberg, Linn ^{[1
,2
]}

von Feilitzen, Kalle ^{[1
,2
]}

Oksvold, Per ^{[1
,2
]}

Lindskog, Cecilia ^{[4
]}

Forsstrom, Bjorn ^{[1
,2
]}

Uhlen, Mathias ^{[1
,2
,5
]}

机构：

[1] KTH Royal Inst Technol, Sci Life Lab, SE-17121 Stockholm, Sweden

[2] KTH Royal Inst Technol, Dept Prot Sci, SE-10691 Stockholm, Sweden

[3] Karolinska Inst, Karolinska Univ Hosp, Dept Oncol Pathol, SE-17177 Stockholm, Sweden

[4] Uppsala Univ, Dept Immunol Genet & Pathol, SE-75185 Uppsala, Sweden

[5] Tech Univ Denmark, Novo Nordisk Fdn Ctr Biosustainabil, DK-2970 Horsholm, Denmark

来源：

NATURE COMMUNICATIONS | 2018年 / 9卷

关键词：

PROTEIN; EXPRESSION;

D O I：

10.1038/s41467-018-06642-y

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：10

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