EMD Millipore
Anti-Stage-Specific Embryonic Antigen-4, clone MC-813-70
MAB4304
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SSEA4 antigen is expressed upon the surface of human embryonic stem (ES) cells and human induced pluripotent stem cells (iPSC). This antibody reacts with the stage-specific embryonic antigen-4 (SSEA-4), a glycolipid carbohydrate epitope that is expressed upon the surface of human teratocarcinoma stem cells (EC), human embryonic germ cells (EG) human ES and human iPSC. Since SSEA-4 is expressed on the surface of cells,it can be used in FACS analysis or immunofluorescence assays without fixing.
Immunocytochemical staining
Human ES/iPS cells
Anti-SSEA4 antibody diluted1:1000 in 3% bovine serum albumin (BSA) in phosphate buffered saline + 0.05% Tween-20 (PBST), incubated overnight in a cold room with rocking.
3% BSA in PBST , for 3 hrs at room temperature with rocking.
Goat anti-mouse Alexa-488 (Life Technologies, cat #A21042), diluted 1:1000 blocking buffer, incubated 2 hrs at room temperature with rocking.
N/A
Confocal microscope (Olympus DSU Spinning Disk Confocal)
Staining appeared on the surface of hES/iPSC and did not overlap with the nucleus marker hoeschst. Staining was pretty strong with no background. Differentiated cells and original fibroblast cells did not show this marker.
None.
Specific staining for human ES and iPS cells.
Nothing.
A good marker for hES/hiPSC. This antibody gives us pretty good results showing pluripotent marker which correlated to mRNA level by RT-PCR as well.