Antibody elution method for multiple immunohistochemistry on primary antibodies raised in the same species and of the same subtype
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences . Biomedical Sciences
New York, N.Y.
The journal of histochemistry and cytochemistry. - New York, N.Y.
, p. 567-575
University of Antwerp
Search: >> Advanced Search Institution: Universiteit Antwerpen Bibliotheek | Sign In via User Name/Password Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact Originally published as JHC exPRESS on February 16, 2009. doi:10.1369/jhc.2009.953240 This Article Full Text Full Text (PDF) Supplemental Data for this article All Versions of this Article: jhc.2009.953240v1 57/6/567 most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Pirici, D. Articles by Stanescu, R. PubMed PubMed Citation Articles by Pirici, D. Articles by Stanescu, R. Social Bookmarking What's this? Journal of Histochemistry and Cytochemistry Volume 57 (6): 567-575, 2009 Copyright ©The Histochemical Society, Inc. Antibody Elution Method for Multiple Immunohistochemistry on Primary Antibodies Raised in the Same Species and of the Same Subtype Daniel Pirici, Laurentiu Mogoanta, Samir Kumar-Singh, Ionica Pirici, Claudiu Margaritescu, Cristina Simionescu and Radu Stanescu Department of Histology (DP,LM) and Department of Pathology (CM,CS), University of Medicine and Pharmacy Craiova, Craiova, Romania; Neurodegenerative Brain Diseases Group, Vlaams Instituut voor Biotechnologie, Department of Molecular Genetics, Laboratory of Neurogenetics, Institute Born Bunge, University of Antwerp, Antwerp, Belgium (DP,SK-S); and Emergency County Hospital 1, Craiova, Romania (IP,RS) Correspondence to: Daniel Pirici, MD, Department of Histology, University of Medicine and Pharmacy Craiova, Petru Rares Street 2, 200349 CraiovaDolj, Romania. E-mail: email@example.com Double or multiple antigen labeling in IHC classically relies on the existence of primary antibodies raised in different species or of different IgG isotypes to ensure the specific labeling with the secondary detection systems. However, suitable pairs of primary antibodies are not always available or the best choice (e.g., as diagnostic tools). During the last few years, several methods have been proposed to overcome this, but none of them offers the flexibility needed for reliable double or multiple enzymatic or fluorescent IHC. We present here a procedure that elutes the antibodies after a first round of immunolabeling, which, in combination with precipitation-based detection systems, allows multiple IHC rounds even for primary antibodies raised in the same species and IgG isotype. Compared with other proposed methods, this procedure ensures a reliable enzymatic or fluorescent staining without cross-reactivity and without loss of tissue antigenicity, thus offering a flexible tool for colocalization studies and pathological diagnosis. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.