Protocol for sorting cancer stem cells using a combination of anti-CD133 antibody and lectin cyanovirin-N

CD133 is widely used as a marker to isolate cancer stem cells (CSCs). However, the structural ambiguity of N-glycan of CD133 limits its application in the isolation of CSCs. Here, we present a protocol to sort CSCs from tumor samples by combining CD133 with α-1,2-high-mannose type glycan chains. We describe steps for purifying and biotinylating cyanovirin-N (CVN) proteins and sorting tumor cells and CD133⁺α-1,2-Man⁺ (α-1,2-linked mannose⁺) cells to provide more specific markers for CSCs. For complete details on the use and execution of this protocol, please refer to Wei et al.¹.