title: The location of (1→3)-β-glucans in the walls of pollen tubes of Nicotiana alata using a (1→3)-β-glucan-specific monoclonal antibody.
abstract: The location of the (1→3)-β-glucan, callose, in the walls of pollen tubes in the style of Nicotiana alata Link et Otto was studied using specific monoclonal antibodies. The antibodies were raised against a laminarinhaemocyanin conjugate. One antibody selected for further characterization was specific for (1→3)-β-glucans and showed no binding activity against either a cellopentaose-bovine serum albumin (BSA) conjugate or a (1→3, 1→4)-β-glucan-BSA conjugate. Binding was inhibited by (1→3)-β-oligoglucosides (DP, 3-6) with maximum competition being shown by laminaripentaose and laminarihexaose, indicating that the epitope included at least five (1→3)-β-linked glucopyranose residues. The monoclonal antibody was determined to have an affinity constant for laminarihexaose of 2.7. 10(4)M(-1). When used with a second-stage gold-labelled, rabbit anti-mouse antibody, the monoclonal antibody probe specifically located the (1→3)-β-glucan in the inner wall layer of thin sections of the N. alata pollen tubes.
authors.affiliation: Centre for Protein and Enzyme Technology and Department of Biochemistry, La Trobe University, 3083, Melbourne, Australia.
authors.forename: P J
authors.initials: PJ
authors.lastname: Meikle
authors.forename: I
authors.initials: I
authors.lastname: Bonig
authors.forename: N J
authors.initials: NJ
authors.lastname: Hoogenraad
authors.forename: A E
authors.initials: AE
authors.lastname: Clarke
authors.forename: B A
authors.initials: BA
authors.lastname: Stone