Rabbit serum immunized with S. cerevisiae deficient in the alpha1->3 mannosyltransferase (deltaMnn1)

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Experiment Name	Rabbit serum immunized with S. cerevisiae deficient in the alpha1->3 mannosyltransferase (deltaMnn1)
Associated Publication	Polysaccharide mimicry of the epitope of the broadly neutralizing anti-HIV antibody, 2G12, induces enhanced antibody responses to self oligomannose glycans. Dunlop DC, Bonomelli C, Mansab F, Vasiljevic S, Doores KJ, Wormald MR, Palma AS, Feizi T, Harvey DJ, Dwek RA, Crispin M, Scanlan CN Glycobiology, 20, 812-23 2010 Jul
Analyte Name	S. cerevisiae deltaMnn1 immune serum
Analyte Family	Antibodies
Analyte Information	Anti-serum elicited after rabbit immunization with Saccharomyces cerevisiae deficient for the alpha->3 mannosyltransferase gene (deltaMnn1), which is responsible for biosynthesis of the variable and polydisperse Manalpha->3Man'cap' found on the nonreducing termini of the Manalpha->2Man branches of Saccharomyces cerevisiae mannan but lacking in mammals. Aim was to focus the immune response toward mammalian glycans.
Concentration	1/25
Protocol	After blocking arrayed slides with 0.02% (v/v) Blocker Casein (Pierce), 1% (w/v) bovine serum albumin (Sigma A8577) in Hepes buffered saline (5 mM Hepes, pH 7.4, 150 mM NaCl, 5 mM CaCl2), S. cerevisiae deltaMnn1 immune serum was assayed at 1/25, followed by biotinylated anti-rabbit IgG (Vector) 1:200. Binding was detected using Alexa Fluor-647-labeled streptavidin from Molecular Probes (1 ug/ml). As diluent the blocker solution was used.
Other Comments	N/A