Family 32 CBM from Cellvibrio mixtus cellulose (CmUnk16A)

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Experiment Name	Family 32 CBM from Cellvibrio mixtus cellulose (CmUnk16A)
Associated Publication	Unravelling glucan recognition systems by glycome microarrays using the designer approach and mass spectrometry. Palma AS, Liu Y, Zhang H, Zhang Y, McCleary BV, Yu G, Huang Q, Guidolin LS, Ciocchini AE, Torosantucci A, Wang D, Carvalho AL, Fontes CM, Mulloy B, Childs RA, Feizi T, Chai W Molecular & cellular proteomics : MCP, 14, 974-88 2015 Apr
Analyte Name	CmCBM32-2
Analyte Family	Carbohydrate-binding modules
Analyte Information	C-terminal sequence region of the Cellvibrio mixtus CmUnk16A modular enzyme gene (lic26A-cel5A) encoding a family 32 CBM, fused with a C-terminal hexa-histidine tag and recombinantly expressed in Escherichia coli. CmUnk16A is a modular containing a GH16 catalytic domain but the role in the hydrolysis of cell wall polysaccharides has not yet been assigned.
Concentration	5 ug/ml
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), CmCBM32-2 was precomplexed with mouse monoclonal anti-poly-histidine and biotinylated anti-mouse IgG antibodies (both from Sigma) in a ratio of 1:3:3 (by weight) and overlaid onto the arrays at 5 ug/ml. 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