Drosophila Notum_conditioned medium

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Experiment Name	Drosophila Notum_conditioned medium
Associated Publication	Notum deacylates Wnt proteins to suppress signalling activity. Kakugawa S, Langton PF, Zebisch M, Howell S, Chang TH, Liu Y, Feizi T, Bineva G, O'Reilly N, Snijders AP, Jones EY, Vincent JP Nature, 519, 187-192 2015 Mar 12
Analyte Name	Drosophila Notum
Analyte Family	Animal lectins
Analyte Information	Conditioned medium obtained from a stable S2 line expressing C-terminal V5-tagged dNotum which was generated by transfection of S2 cells with pActin -Notum-V5 and pCoHygro followed by drug selection. Notum is a highly conserved secreted feedback inhibitor of Wnt signalling.
Concentration	Neat
Protocol	After blocking arrayed slides with 3% (w/v) bovine serum albumin (Sigma A8577) in Hepes buffered saline (5 mM Hepes, pH 7.4, 150 mM NaCl, 5 mM CaCl2), conditioned medium obtained from S2 cells expressing V5-tagged dNotum was overlaid onto the arrays as neat (undiluted) followed by of anti-V5 mouse monoclonal antibody (Invitrogen), 5 ug/ml, and then by biotinylated anti-mouse IgG (Sigma), 1:200. Binding was detected using Alexa Fluor-647-labeled streptavidin from Molecular Probes (1 ug/ml).
Other Comments	No binding was observed when control medium was used instead of the conditioned medium or when the anti-V5 was used in the absence of the dNotum medium.