Immunofluorescent analysis of SSEA-3 using anti-SSEA-3 monoclonal antibody (Product # MA1-020) shows expression in human teratocarcinoma NCCIT cells (shown in green) but not in negative control HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a rat monoclonal antibody recognizing SSEA-3 (Product # MA1-020), at a dilution of 1:50 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with a goat-anti-rat IgM secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rat / IgM|
|Immunogen||Fischer rat immunized with 4-8 cell stage mouse embryos|
|Purification||Affinity chromatography - MBP|
|Storage buffer||potassium phosphate with 0.15M NaCl, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100|
|Immunofluorescence (IF)||1:100 - 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.
MA1-020 has been successfully used in immunofluorescence and flow cytometry applications on human samples.
SSEA-3 (stage-specific embryonic antigen 3) is a cell surface antigen, expressed along with SSEA-4, TRA-1-60 and TRA-1-81 in embryonic stem cells, embryonal carcinoma cells, and induced pluripotent stem cells (iPS). In human cells, these surface markers are down-regulated during the differentiation process, while SSEA1 is absent in undifferentiated human stem cells and upregulated on the cell surface after retinoic acid mediated differentiation. Conversely, in mouse stem cells, SSEA-1 expression decreases with differentiation while SSEA-3 and SSEA-4 expression increase.
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