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Immunofluorescent analysis of SSEA-5 (green) on NTERA-2 embryonal carcinoma cells. Formalin fixed cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) a SSEA-5 monoclonal antibody (Product # MA1-144) at a dilution of 1:400 for 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|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|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50|
|Immunofluorescence (IF)||1:50 - 1:400|
* 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 shows positive staining of SSEA-5 positive pluripotent cells within cultured human embryonal carcinoma NCCIT and NTERA-2 cells. Mouse teratocarcinoma cell line F9 was used as a negative control and showed no staining.
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.