Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Antibody Validation Strategy for Nuclear Receptors.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science. KTH, Centres, Science for Life Laboratory, SciLifeLab. (Cecilia Williams)ORCID iD: 0000-0002-0602-2062
2019 (English)In: Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029, Vol. 1966, p. 79-99Article in journal (Refereed) Published
Abstract [en]

Antibodies are invaluable biological tools that we can use to detect the presence, location, or alteration of nuclear receptors. However, antibodies frequently cross-react with other proteins and their performance can vary from batch to batch, from application to application and from lab to lab. When each lot of antibody is not thoroughly validated for each assay, each sample type, and each lab and user, antibody-based assays can lead to flawed interpretations and reproducibility problems. In this chapter, we describe a scheme for thorough antibody validation, suitable for nuclear receptors. The method is based on using highly characterized positive and negative controls assembled into a validation tissue microarray (TMA). Through correlation of immunohistochemical staining (IHC) and mRNA levels over multiple tissues, use of current public databases, and assessment of binding to intended and nonintended targets using western blotting (WB), immunoprecipitation (IP), and mass spectrometry (MS), we describe a path for thoroughly validation of antibodies.

Place, publisher, year, edition, pages
2019. Vol. 1966, p. 79-99
Keywords [en]
Antibody, Estrogen receptor, Immunohistochemistry, Immunoprecipitation, Mass spectrometry, Nuclear receptors, RNA-Seq, Tissue microarray, Validation, Western blotting
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:kth:diva-257973DOI: 10.1007/978-1-4939-9195-2_7PubMedID: 31041740Scopus ID: 2-s2.0-85065404524OAI: oai:DiVA.org:kth-257973DiVA, id: diva2:1349600
Note

QC 20190917

Available from: 2019-09-09 Created: 2019-09-09 Last updated: 2019-09-17Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textPubMedScopus

Authority records BETA

Williams, Cecilia

Search in DiVA

By author/editor
Williams, Cecilia
By organisation
Protein ScienceScience for Life Laboratory, SciLifeLab
In the same journal
Methods in Molecular Biology
Biomedical Laboratory Science/Technology

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 8 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf