Galectin-6(GAL6) is a protein that in humans is encoded by the LGALS6 gene. The galectins constitute a large family of carbohydrate-binding proteins with specificity for N-acetyl-lactosamine-containing glycoproteins. At least 14 mammalian galectins, which share structural similarities in their carbohydrate recognition domains (CRD), have been identified. Galectin-6 may also be involved in cell-cell and cell-matrix interactions and exogenous galectin has been found to accelerate the re-epithelialization of wounds. It can agglutinate red blood. In this case, we chose rabbit erythrocyte (RaE) to assay its ability of agglutination. A general procedure for hemagglutination assay (or haemagglutination assay; HA) is as follows, two-fold dilute the recombinant Mu GAL6 with 0.9% sodium chloride injection, add 50μL a serial dilution of GAL6 to each well of a U or V-bottom shaped 96-well microtiter plate. The final well serves as a negative control with no GAL6, replace with 50μL 0.9% sodium chloride injection. Then add 50μL 1% rabbit erythrocyte to each well and mixed gently. The plate is incubated for 3 hours at room temperature. The results are shown in Figure 1. It was obvious that the minimal effective concentration of GAL6 is 3.125 μg/mL.

Figure 2. The hemagglutination assay of GAL6 in V- bottom shaped 96-well microtiter plate.