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The risk of using monoclonal or polyclonal commercial antibodies: controversial results on porcine sperm CD44 receptor identification
Linköping University, Department of Clinical and Experimental Medicine, Division of Children's and Women's health. Linköping University, Faculty of Medicine and Health Sciences. Univ Autonoma Barcelona, Spain.
Univ Autonoma Barcelona, Spain.
Linköping University, Department of Clinical and Experimental Medicine, Division of Children's and Women's health. Linköping University, Faculty of Medicine and Health Sciences.ORCID iD: 0000-0002-5194-2124
2019 (English)In: Reproduction in domestic animals, ISSN 0936-6768, E-ISSN 1439-0531, Vol. 54, no 4, p. 733-737Article in journal (Refereed) Published
Abstract [en]

Presence of the hyaluronan (hyaluronic acid, HA) receptor CD44 on spermatozoa has been difficult to pursue, mostly obeying to the use of different commercial mono- and/or polyclonal antibodies, often lacking proper controls. Here, we describe how the presence (Western blotting) and specific location (immunocytochemistry) of the CD44 receptor differs in ejaculated pig spermatozoa depending on the type of antibody and protocol used. While we were able to detect binding to spermatozoa and mark its presence in the sperm membrane, the use of blocking peptides clearly indicated that only the monoclonal antibody could confirm the specific presence and location of the CD44 receptor, whereas the polyclonal antibody was detecting multiple presumed CD44 isoforms or degraded proteins thus proving unspecific. These results call for strict protocols when attempting immunological determination of sperm membrane receptors.

Place, publisher, year, edition, pages
WILEY , 2019. Vol. 54, no 4, p. 733-737
Keywords [en]
CD44 receptor; hyaluronan; monoclonal antibody; pig; polyclonal antibody; spermatozoa
National Category
Medicinal Chemistry
Identifiers
URN: urn:nbn:se:liu:diva-157233DOI: 10.1111/rda.13415ISI: 000466408600010PubMedID: 30746820OAI: oai:DiVA.org:liu-157233DiVA, id: diva2:1324210
Note

Funding Agencies|Lions Forskningsfond 2016-postdoc, Linkoping, Sweden [DNR LIU-2016-00641]; Forskningsradet i Sydostra Sverige [473121, 745971]; Svenska Forskningsradet Formas [2017-00946]

Available from: 2019-06-13 Created: 2019-06-13 Last updated: 2019-06-13

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Alvarez-Rodriguez, ManuelRodriguez-Martinez, Heriberto
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