Alcian blue and silver staining

The Clec10a-L was resolved by SDS–polyacrylamide gel electrophoresis (SDS-PAGE). Immediately after electrophoresis, the gels were washed with deionized water and 10% acetic acid buffer (10% acetic acid and 30% ethanol in deionized water) and stained or not with Alcian Blue solution (pH 2.5) (catalog no. 013-13801, Wako) at room temperature for 2 hours, followed by destaining with 3% acetic acid buffer (3% acetic acid in deionized water) and 10% acetic acid buffer. The gels were then stained with silver according to the manufacturer’s instructions (Pierce Silver Stain Kit, catalog no. 24612, Thermo Fisher Scientific).