Internal family 11 CBM from Clostridium thermocellum Lic26A-Cel5E endoglucanase

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Experiment Name	Internal family 11 CBM from Clostridium thermocellum Lic26A-Cel5E endoglucanase
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	CtCBM11
Analyte Family	Carbohydrate-binding modules
Analyte Information	Internal sequence region of the Clostridium thermocellum Lic26A-Cel5A modular enzyme gene (lic26A-cel5A) encoding the family 11 CBM, fused with a C-terminal hexa-histidine tag and recombinantly expressed in Escherichia coli. Lic26A-Cel5E modular enzyme contains GH5 and GH26 catalytic domains that were shown to display beta1,4 and beta-1,3 - 1,4 mixed linked endoglucanase activity, respectively.
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), CtCBM11 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