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Flow cytometry analysis of SSEA-5 in NCCIT (red filled histogram) and HeLa cells (red unfilled histogram) compared to unstained cells (blue histogram). Positive staining is observed on NCCIT cells when compared to no antibody control (middle panel) and to SSEA-5-negative HeLa cells (right panel). Cells were fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with 2% BSA-PBS for 30 minutes at room temperature and incubated with a PE-conjugated SSEA-5 monoclonal antibody (Product # MA1-144-PE) in 2% BSA-PBS at a dilution of 1:100 for 60 minutes at room temperature. Cells were washed and re-suspended in PBS for FACS analysis.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Undifferentiated H9 human embryonic stem cells|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50-1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Immunofluorescence and flow cytometric analysis of MA1-144-PE shows positive staining of SSEA-5 in human embryonal carcinoma NCCIT cells.
Stage-Specific Embryonic Antigen-5 (SSEA-5) is an oligosaccharide that can be part of glycosphingolipids and glycoproteins. It contains the motif Fuc1-2Gal-beta1-3GlcNAc-beta, which is characteristic of H-1 antigens. SSEA-5 is found in the inner cell mass of the human blastocyst during embryogenesis, on undifferentiated human embryonic stem cells and on the surface of induced pluripotent cells. SSEA-5 was found to proceed the loss of other known stem cell markers and the combination of SSEA-5 with either CD9 and CD90 or CD50 and CD200 was superior in identifying and eliminating rare undifferentiated human ES cells from differentiated cell populations than the use of other established stem cell markers.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.