» Articles » PMID: 32852223

Antibody Validation for Protein Expression on Tissue Slides: a Protocol for Immunohistochemistry

Overview
Journal Biotechniques
Date 2020 Aug 28
PMID 32852223
Citations 27
Authors
Affiliations
Soon will be listed here.
Abstract

Antibodies play a crucial role in basic research and clinical decision-making. However, there are no standardized algorithms or guidelines to ensure their accuracy and validity. There have been efforts to generate consensus, but, with the exception of clinical labs, antibody validation remains variable in the literature and sometimes in clinical practice. Here we focus on immunohistochemistry, an example of a scientific and clinical tool where validation of antibodies is critical. We describe a protocol that we use to validate antibodies specifically for immunohistochemistry, including some of the pillars of antibody validation from Uhlen 2016, as an example of a rigorous approach to build antibody-based tests for both basic and translational science labs and for the clinic.

Citing Articles

Assessing expression patterns of PTGR1, a potential biomarker for acylfulven sensitivity in urothelial carcinoma.

Stormoen D, Lehn S, Mouw K, Szallasi Z, Melchior L, Dohn L Sci Rep. 2024; 14(1):27448.

PMID: 39523403 PMC: 11551215. DOI: 10.1038/s41598-024-79334-x.


BCAM (basal cell adhesion molecule) protein expression in different tumor populations.

Burela S, He M, Trontzas I, Gavrielatou N, Schalper K, Langermann S Discov Oncol. 2024; 15(1):381.

PMID: 39207605 PMC: 11362396. DOI: 10.1007/s12672-024-01244-1.


Quantitative Measurement of HER2 Expression in Non-Small Cell Lung Cancer With a High-Sensitivity Assay.

Liu M, Vathiotis I, Robbins C, Chan N, Moutafi M, Burela S Mod Pathol. 2024; 37(9):100556.

PMID: 38964502 PMC: 11416319. DOI: 10.1016/j.modpat.2024.100556.


Validation of an organ mapping antibody panel for cyclical immunofluorescence microscopy on normal human kidneys.

Brewer M, Migas L, Clouthier K, Allen J, Anderson D, Pingry E Am J Physiol Renal Physiol. 2024; 327(1):F91-F102.

PMID: 38721662 PMC: 11390132. DOI: 10.1152/ajprenal.00426.2023.


Detecting Human Epidermal Growth Factor Receptor 2 (HER2) Amplification: Proof of Concept of an Alternative Approach.

Mudgal S, Paul P, Ravi B, Agrawal S, Kalra A, Rao S Cureus. 2023; 15(9):e44785.

PMID: 37809181 PMC: 10558136. DOI: 10.7759/cureus.44785.


References
1.
Bradbury A, Pluckthun A . Getting to reproducible antibodies: the rationale for sequenced recombinant characterized reagents. Protein Eng Des Sel. 2015; 28(10):303-5. DOI: 10.1093/protein/gzv051. View

2.
Sfanos K, Yegnasubramanian S, Nelson W, Lotan T, Kulac I, Hicks J . If this is true, what does it imply? How end-user antibody validation facilitates insights into biology and disease. Asian J Urol. 2019; 6(1):10-25. PMC: 6363603. DOI: 10.1016/j.ajur.2018.11.006. View

3.
Andersson S, Sundberg M, Pristovsek N, Ibrahim A, Jonsson P, Katona B . Insufficient antibody validation challenges oestrogen receptor beta research. Nat Commun. 2017; 8:15840. PMC: 5501969. DOI: 10.1038/ncomms15840. View

4.
Gustavson M, Rimm D, Dolled-Filhart M . Tissue microarrays: leaping the gap between research and clinical adoption. Per Med. 2018; 10(5):441-451. DOI: 10.2217/pme.13.42. View

5.
Bandrowski A, Brush M, Grethe J, Haendel M, Kennedy D, Hill S . The Resource Identification Initiative: A Cultural Shift in Publishing. Neuroinformatics. 2015; 14(2):169-82. PMC: 5072392. DOI: 10.1007/s12021-015-9284-3. View