SSEA-4 is expressed on the surface of human embryonal carcinoma (EC) cells (the pluripotent stem cells of teratocarcinomas), human embryonic germ cells (EG), and human embryonic stem cells (ES). Expression of SSEA-4 is down-regulated following differentiation of human EC cells. In contrast, the differentiation of murine EC and ES cells may be accompanied by an increase in SSEA-4 expression.1-4
Produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with the human embryonal carcinoma cell line 2102 Ep.
Flow Cytometry, Immunohistochemistry
Immunohistochemistry: This antibody has been used with the appropriate secondary reagents at a concentration of 5-10ug/mL in fixed cells. Cells were fixed with 4% paraformaldehyde in PBS at room temperature for 20 minutes, then blocked with PBS containing 10% normal donkey serum and 1% BSA at room temperature for 45 minutes. After blocking, cells were incubated with diluted primary antibody overnight at 4 degrees C followed by Rhodamine Red(TM) coupled anti-mouse IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. This antibody can also be used in unfixed, shock frozen tissue at a concentration of 10ug/mL.
Flow Cytometry: Dilute this antibody to 0.1 mg/mL and add 5ul of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200ul. The binding of unlabeled monoclonal antibodies may be visualized by adding 10ul of a 25ug/mL stock solution of a secondary developing reagent such as goat anti-mouse IgG conjugated to a fluorochrome.
Optimal dilutions to be determined by the researcher.
Storage and Stability:
Lyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.