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. 2022 Mar 31;55(3):142–147. doi: 10.5483/BMBRep.2022.55.3.090

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Copyright © 2022 by the The Korean Society for Biochemistry and Molecular Biology

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 1 — Monoclonal antibody K312 specifically binds to human pluripotent stem cells (PSCs). (A) Flow cytometry analysis of the expression of PSC surface markers and the target of K312 on H9 human embryonic stem cells (H9 hESCs). (B) K312 specifically binds to human pluripotent cell lines (iPS-fAD, iPS-SPD, NTERA-2, and NCCIT), but not to J1 mouse embryonic stem cells. (C) Flow cytometry analysis of the correlative expression of PSC markers and the target of K312. H9 hESCs were stained with K312 and the indicated PSC marker-specific antibodies. (D) Immunofluorescence staining shows the co-localization of E-cadherin and the target protein of K312 on H9 hESCs. Scale bar: 25 μm. (E) Immunoprecipitation of H9 hESC lysates with control mouse IgG (mIgG1) or K312; cells were analyzed by immunoblotting. Black arrow indicates a protein band of approximately 60 kDa appearing after immunoprecipitation with K312. (F) Deglycosylation assay. H9 hESC or NCCIT lysates were treated with PNGase F or left untreated for the indicated times, and analyzed by immunoblotting using K312. Cell lysates not treated with PNGase F were used as control (Ctrl).