rBC2LCN, a Novel Lectin Probe for Human Pluripotent Stem Cells
The human pluripotent stem cell (hPSC)-specific lectin rBC2LCN (recombinant N-terminal domain of BC2L-C from Burkholderia cenocepacia) was identified by comprehensive glycome analysis of a large number of hPSCs with the aid of a high-density lectin microarray. rBC2LCN is a 16 kDa protein that recognizes O-glycans containing H type-3 structure (Fucα1-2Galβ1-3GalNAc), which are heavily expressed on podocalyxin, a member of the sialomucin family. rBC2LCN has been found to be a practical hPSC-specific probe that can be used to stain formaldehyde-fixed hPSCs. It can also be supplemented into cell culture media to enable the live staining of hPSCs with no apparent toxicity. Unlike conventional antibodies, which require mammalian cells for their production, rBC2LCN can be easily produced in large amounts and in a cost-effective manner in an E. coli expression system. Based on these strengths, rBC2LCN has now been made commercially available as a novel reagent for the detection of hPSCs. Here, experimental protocols for the fluorescence staining of fixed and live hPSCs and flow cytometry using rBC2LCN are described.
KeywordsPluripotent stem cells Lectin Staining Flow cytometry O-glycan H type-3
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