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Antibody Validation...
Antibody Validation for Estrogen Receptor Beta
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- Birgersson, Madeleine (författare)
- Karolinska Institutet,KTH,Cellulär och klinisk proteomik,Science for Life Laboratory, SciLifeLab,Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden,Williams
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- Katona, Borbala (författare)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Institutionen för immunologi, genetik och patologi
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- Lindskog, Cecilia (författare)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Cancerprecisionsmedicin
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- Pontén, Fredrik (författare)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Cancerprecisionsmedicin
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- Williams, Cecilia, Professor, 1969- (författare)
- Karolinska Institutet,KTH,Cellulär och klinisk proteomik,Science for Life Laboratory, SciLifeLab,Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden,Cecilia Williams
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(creator_code:org_t)
- 2022-02-04
- 2022
- Engelska.
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Ingår i: Methods in Molecular Biology. - New York, NY : Springer Nature. - 1064-3745 .- 1940-6029. ; 2418, s. 1-23, s. 1-23
- 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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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- Antibodies can cross-react with proteins other than their intended targets, and antibody-based applications can, if not properly validated, lead to flawed interpretations. When evaluating 13 anti-estrogen receptor beta (ERβ) antibodies in 2017, we concluded that only one of them was specific. Applying this antibody in immunohistochemistry of over 44 different normal human tissues and 20 types of cancers revealed ERβ expression in only a few selected tissues. This aligned with mRNA evidence but contradicted a large set of published literature. ERβ protein expression continues to be reported in tissues without clear support by mRNA expression. In this chapter, we describe how ERβ antibodies can be thoroughly validated and discuss selection of well-characterized positive and negative controls. The validation scheme presented is applicable for immunohistochemistry and Western blotting. The protocol includes evaluation of mRNA evidence, use of public databases, assessment of on- and off-target binding, and an optional step for corroboration with immunoprecipitation and mass spectrometry.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Annan medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Other Medical Biotechnology (hsv//eng)
Nyckelord
- Antibody
- Estrogen receptor beta
- Immunohistochemistry
- RNA-Seq
- Validation
- Western blotting
- Bioteknologi
- Biotechnology
- Biology with specialization in Molecular Biology
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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