Pre-hybridisation: an efficient way of suppressing endogenous biotin-binding activity inherent to biotin–streptavidin detection system

2.50
Hdl Handle:
http://hdl.handle.net/10547/560960
Title:
Pre-hybridisation: an efficient way of suppressing endogenous biotin-binding activity inherent to biotin–streptavidin detection system
Authors:
Ahmed, Raju; Spikings, Emma ( 0000-0001-8387-2098 ) ; Zhou, Shaobo ( 0000-0001-5214-2973 ) ; Thompsett, Andrew; Zhang, Tiantian
Abstract:
Endogenous biotin or biotinylated protein binding activity is a major drawback to biotin-avidin/streptavidin detection system. The avidin/streptavidin conjugate used to detect the complex of the biotinylated secondary antibody and the primary antibody binds to endogenous biotin or biotinylated proteins leading to non-specific signals. In Western blot, the endogenous biotin or biotinylated protein binding activity is usually manifested in the form of ~72kDa, ~75kDa and ~150kDa protein bands, which often mask the signals of interest. To overcome this problem, a method based on prior hybridisation of the biotinylated secondary antibody and the streptavidin conjugate was developed. The method was tested alongside the conventional biotin-streptavidin method on proteins extracted from zebrafish (Danio rerio) embryos. Results showed that the newly developed method efficiently suppresses the endogenous biotin or biotinylated protein binding activity inherent to the biotin-streptavidin detection system.
Affiliation:
University of Bedfordshire; University of East London; Bournemouth University
Citation:
Ahmed, R., Spikings, E., Zhou, S.B., Thompsett, A., Zhang, T.T. (2014) 'Pre-hybridisation: an efficient way of suppressing endogenous biotin-binding activity inherent to biotin–streptavidin detection system', Journal of Immunological Methods, 406, pp.143 - 147.
Publisher:
Elsevier
Journal:
Journal of Immunological Methods
Issue Date:
Apr-2014
URI:
http://hdl.handle.net/10547/560960
DOI:
10.1016/j.jim.2014.03.010
Additional Links:
http://www.sciencedirect.com/science/article/pii/S0022175914000842
Type:
Article
Language:
en
ISSN:
0022-1759
Appears in Collections:
Biomedical Science

Full metadata record

DC FieldValue Language
dc.contributor.authorAhmed, Rajuen
dc.contributor.authorSpikings, Emmaen
dc.contributor.authorZhou, Shaoboen
dc.contributor.authorThompsett, Andrewen
dc.contributor.authorZhang, Tiantianen
dc.date.accessioned2015-07-24T09:43:14Zen
dc.date.available2015-07-24T09:43:14Zen
dc.date.issued2014-04en
dc.identifier.citationAhmed, R., Spikings, E., Zhou, S.B., Thompsett, A., Zhang, T.T. (2014) 'Pre-hybridisation: an efficient way of suppressing endogenous biotin-binding activity inherent to biotin–streptavidin detection system', Journal of Immunological Methods, 406, pp.143 - 147.en
dc.identifier.issn0022-1759en
dc.identifier.doi10.1016/j.jim.2014.03.010en
dc.identifier.urihttp://hdl.handle.net/10547/560960en
dc.description.abstractEndogenous biotin or biotinylated protein binding activity is a major drawback to biotin-avidin/streptavidin detection system. The avidin/streptavidin conjugate used to detect the complex of the biotinylated secondary antibody and the primary antibody binds to endogenous biotin or biotinylated proteins leading to non-specific signals. In Western blot, the endogenous biotin or biotinylated protein binding activity is usually manifested in the form of ~72kDa, ~75kDa and ~150kDa protein bands, which often mask the signals of interest. To overcome this problem, a method based on prior hybridisation of the biotinylated secondary antibody and the streptavidin conjugate was developed. The method was tested alongside the conventional biotin-streptavidin method on proteins extracted from zebrafish (Danio rerio) embryos. Results showed that the newly developed method efficiently suppresses the endogenous biotin or biotinylated protein binding activity inherent to the biotin-streptavidin detection system.en
dc.language.isoenen
dc.publisherElsevieren
dc.relation.urlhttp://www.sciencedirect.com/science/article/pii/S0022175914000842en
dc.rightsArchived with thanks to Journal of Immunological Methodsen
dc.subjectbiotinylated secondary antibodyen
dc.subjectendogenous biotin bindingen
dc.subjectnon-specific bandsen
dc.subjectQdot 625 streptavidin conjugateen
dc.subjectStreptavidinen
dc.subjectWestern bloten
dc.titlePre-hybridisation: an efficient way of suppressing endogenous biotin-binding activity inherent to biotin–streptavidin detection systemen
dc.typeArticleen
dc.contributor.departmentUniversity of Bedfordshireen
dc.contributor.departmentUniversity of East Londonen
dc.contributor.departmentBournemouth Universityen
dc.identifier.journalJournal of Immunological Methodsen
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