Alexa647 linked anti mouse igg

Recombinant trimeric HA0 was expressed in HEK293S GnTI^-/- (American Type Culture Collection), purified and analyzed on glycan array as previously described [35 (link)]. Briefly, soluble trimeric HA (50 μg mL^-1) was pre-complexed with the anti-HIS mouse antibody (MA1-21315, Thermo Fisher Scientific, Waltham, MA) and the Alexa647-linked anti-mouse IgG (A-21235, Thermo Fisher Scientific) at 4:2:1 molar ratio for 15 min on ice in 50 μL PBST. This complex was incubated on the array surface in a humidified chamber for 60 min before washing and analysis using an Innoscan 1100AL microarray scanner (Innopsys, Chicago, IL). Fluorescent signal intensity was measured using Mapix (Innopsys) and mean intensity minus mean background of 4 replicate spots was calculated. A complete list of the glycans on the array is presented in S11 Fig.

Recombinant trimeric HA0 was expressed in HEK 293S GnTI^-/- cells, purified and analyzed on a glycan array as previously described (Peng et al., 2017 (link)). Briefly, soluble trimeric HA (50 μg mL^-1) was pre-complexed with the anti-HIS mouse antibody (Thermo Fisher Scientific) and the Alexa647-linked anti-mouse IgG (Thermo Fisher Scientific) at 4:2:1 molar ratio for 15 minutes on ice in 50 μL PBST. This complex was incubated on the array surface in a humidified chamber for 60 minutes before washing and analysis using an Innoscan 1100AL microarray scanner (Innopsys, Chicago, IL). Fluorescent signal intensity was measured using Mapix (Innopsys) and mean intensity minus mean background of 4 replicate spots was calculated. A complete list of the glycans on the array is presented in Data S1.