SDS-PAGE (NBP1-30277)

1. Mix equal volumes of human blood and Alsever's solution (pH 7.0).
(Alsever's solution: NaCl 0.42 g, Sodium citric acid 0.8g, Citric acid 0.055 g, D-glucose 2.05g in DW100 ml)

2. Centrifuge at 15000rpm for 10 minutes and wash four times with PBS.

3. Dilute packed cells in a 0.5 mg/ml trypsin-EDTA solution to give 4% red cell suspension.

4. Incubate for 1h at 37C and wash four times with PBS.

5. Dilute packed cells in PBS to give 4% red cell suspension.

6. Load 50ul of 0.5%BSA-in-0.15M-NaCl solution and 25ul of 4%-Red-Cell-in-PBS in u shaped wells.

7. Add 25ul of serial diluted galectin protein in PBS to each well plate. (Round bottom 96 well plate)

8. Incubate for 30min at room temperature to observe visible agglutination.