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Antibody Validation...
Antibody Validation in Bioimaging Applications Based on Endogenous Expression of Tagged Proteins
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- Skogs, Marie (författare)
- KTH,Science for Life Laboratory, SciLifeLab
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- Stadler, Charlotte (författare)
- KTH,Science for Life Laboratory, SciLifeLab
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- Schutten, Rutger (författare)
- KTH,Science for Life Laboratory, SciLifeLab
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- Hjelmare, Martin (författare)
- KTH,Science for Life Laboratory, SciLifeLab
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- Gnann, Christian (författare)
- KTH,Science for Life Laboratory, SciLifeLab
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- Björk, Lars (författare)
- Karolinska Institutet,KTH,Science for Life Laboratory, SciLifeLab
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Poser, Ina (författare)
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Hyman, Anthony (författare)
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- Uhlén, Mathias (författare)
- KTH,Science for Life Laboratory, SciLifeLab
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- Lundberg, Emma (författare)
- KTH,Science for Life Laboratory, SciLifeLab
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(creator_code:org_t)
- 2016-10-26
- 2017
- Engelska.
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Ingår i: Journal of Proteome Research. - : American Chemical Society (ACS). - 1535-3893 .- 1535-3907. ; 16:1, s. 147-155
- Relaterad länk:
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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
Ämnesord
Stäng
- 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.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology (hsv//eng)
Nyckelord
- antibody validation
- spatial proteomics
- GFP
- Human Protein Atlas
- Cell Atlas
- subcellular localization
- immunofluorescence
- confocal microscopy
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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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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visa fler...
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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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visa färre...
- Om ämnet
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- MEDICIN OCH HÄLSOVETENSKAP
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MEDICIN OCH HÄLS ...
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och Medicinsk biotek ...
- Artiklar i publikationen
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Journal of Prote ...
- Av lärosätet
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Kungliga Tekniska Högskolan
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Karolinska Institutet