Imunofluorescent analysis of SSEA5 (green) and SSEA4 (red) in human episomal iPSC cells. Cells were grown on vitronectin coated plates, fixed with 4% paraformaldehyde for 15 minutes, and blocked with 3% Blocker BSA (Product # 37525) for 30 minutes at room temperature. Cells were co-stained with DyLight 488-conjugated SSEA5 and DyLight 650-conjugated SSEA4 monoclonal antibodies (top left panel, Product # MA1-144-D488 and MA1-021-D650) or DyLight 488-conjugated Mouse IgG1 and DyLight 650-conjugated Mouse IgG3 isotype control antibodies (right panels, Product # MA1-191-D488 and MA1-190-D650), at a dilution of 1:50 and incubated for 1 hour at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on an EVOS FL Imaging system at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Undifferentiated H9 human embryonic stem cells|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% 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-D488 shows positive staining of SSEA-5 positive pluripotent cells and cultured 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.