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Antibody Validation...
Antibody Validation in Bioimaging Applications Based on Endogenous Expression of Tagged Proteins
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- Skogs, Marie (author)
- KTH,Science for Life Laboratory, SciLifeLab
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- Stadler, Charlotte (author)
- KTH,Science for Life Laboratory, SciLifeLab
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- Schutten, Rutger (author)
- KTH,Science for Life Laboratory, SciLifeLab
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- Hjelmare, Martin (author)
- KTH,Science for Life Laboratory, SciLifeLab
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- Gnann, Christian (author)
- KTH,Science for Life Laboratory, SciLifeLab
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- Björk, Lars (author)
- Karolinska Institutet,KTH,Science for Life Laboratory, SciLifeLab
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Poser, Ina (author)
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Hyman, Anthony (author)
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- Uhlén, Mathias (author)
- KTH,Science for Life Laboratory, SciLifeLab
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- Lundberg, Emma (author)
- KTH,Science for Life Laboratory, SciLifeLab
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(creator_code:org_t)
- 2016-10-26
- 2017
- English.
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In: Journal of Proteome Research. - : American Chemical Society (ACS). - 1535-3893 .- 1535-3907. ; 16:1, s. 147-155
- Related links:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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http://kipublication...
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Abstract
Subject headings
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- Antibodies are indispensible research tools, yet the scientific community has not adopted standardized procedures to validate their specificity. Here we present a strategy to systematically validate antibodies for immunofluorescence (IF) applications using gene tagging. We have assessed the on- and off-target binding capabilities of 197 antibodies using 108 cell lines expressing EGFP-tagged target proteins at endogenous levels. Furthermore, we assessed batch-to-batch effects for 35 target proteins, showing that both the on- and off-target binding patterns vary significantly between antibody batches and that the proposed strategy serves as a reliable procedure for ensuring reproducibility upon production of new antibody batches. In summary, we present a systematic scheme for antibody validation in IF applications using endogenous expression of tagged proteins. This is an important step toward a reproducible approach for context- and application-specific antibody validation and improved reliability of antibody-based experiments and research data.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology (hsv//eng)
Keyword
- antibody validation
- spatial proteomics
- GFP
- Human Protein Atlas
- Cell Atlas
- subcellular localization
- immunofluorescence
- confocal microscopy
Publication and Content Type
- ref (subject category)
- art (subject category)
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To the university's database
- By the author/editor
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Skogs, Marie
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Stadler, Charlot ...
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Schutten, Rutger
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Hjelmare, Martin
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Gnann, Christian
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Björk, Lars
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show more...
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Poser, Ina
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Hyman, Anthony
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Uhlén, Mathias
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Lundberg, Emma
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show less...
- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Medical Biotechn ...
- Articles in the publication
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Journal of Prote ...
- By the university
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Royal Institute of Technology
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Karolinska Institutet