Skip to main content
. 2016 Sep 28;6:34195. doi: 10.1038/srep34195

Figure 4. Detection of core fucosylation by lectin blot and lectin-based ELISA.

Figure 4

(a) Biotinylated SL2-1 (0.6 μg/mL) was used for lectin blotting with different glycosylated and non-glycosylated proteins: 2 μg of mouse IgG (lane 1); 1 μg of mouse IgG (lane 2); 2 μg of mouse IgG treated with PNGase F (lane 3); 2 μg of mouse IgG treated with Endo S (lane 4); 1.5 μg of native CIP (lane 5); 1.5 μg of recombinant CIP expressed in yeast (lane 6); 2 μg of BSA (lane 7); 2 μg of horseradish peroxidase (lane 8). After incubation with the lectin, the membrane was incubated with an HRP-conjugated anti-biotin antibody, and targets visualized using chemiluminescent substrate. (b) Titration of murine IgG and detection by SL2-1 lectin blotting. (c,d) SL2-1 was tested in a direct lectin ELISA assay that utilized a microtiter plate with immobilized IgG and fluorescent dye-labeled lectin (c) or biotin-labeled lectin together with HRP conjugated biotin antibody and TMB colorimetric substrate (d) to quantify core fucosylation of IgG molecules.