Change search
ReferencesLink to record
Permanent link

Direct link
Antibodies Biotinylated Using a Synthetic Z-domain from Protein A Provide Stringent In Situ Protein Detection
KTH, School of Biotechnology (BIO), Protein Technology.
Show others and affiliations
2013 (English)In: Journal of Histochemistry and Cytochemistry, ISSN 0022-1554, E-ISSN 1551-5044, Vol. 61, no 11, 773-784 p.Article in journal (Refereed) Published
Abstract [en]

Antibody-based protein profiling on a global scale using immunohistochemistry constitutes an emerging strategy for mapping of the human proteome, which is crucial for an increased understanding of biological processes in the cell. Immunohistochemistry is often performed indirectly using secondary antibodies for detection, with the benefit of signal amplification. Direct immunohistochemistry instead brings the advantage of multiplexing; however, it requires labeling of the primary antibody. Many antibody-labeling kits do not specifically target IgG and may therefore cause labeling of stabilizing proteins present in the antibody solution. A new conjugation method has been developed that utilizes a modified Z-domain of protein A (ZBPA) to specifically target the Fc part of antibodies. The aim of the present study was to compare the ZBPA conjugation method and a commercially available labeling kit, Lightning-Link, for in situ protein detection. Fourteen antibodies were biotinylated with each method and stained using immunohistochemistry. For all antibodies tested, ZBPA biotinylation resulted in distinct immunoreactivity without off-target staining, regardless of the presence of stabilizing proteins in the buffer, whereas the majority of the Lightning-Link biotinylated antibodies displayed a characteristic pattern of nonspecific staining. We conclude that biotinylated ZBPA domain provides a stringent method for antibody biotinylation, advantageous for in situ protein detection in tissues.

Place, publisher, year, edition, pages
2013. Vol. 61, no 11, 773-784 p.
Keyword [en]
antibody, biotin, conjugation, protein detection, tissue microarray
National Category
Cell Biology
URN: urn:nbn:se:kth:diva-133966DOI: 10.1369/0022155413502360ISI: 000326066300001ScopusID: 2-s2.0-84887331187OAI: diva2:664430
Knut and Alice Wallenberg Foundation

QC 20131115

Available from: 2013-11-15 Created: 2013-11-14 Last updated: 2013-11-15Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textScopus

Search in DiVA

By author/editor
Konrad, AnnaHober, Sophia
By organisation
Protein Technology
In the same journal
Journal of Histochemistry and Cytochemistry
Cell Biology

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 34 hits
ReferencesLink to record
Permanent link

Direct link