TROP-2, also named tumor-associated calcium signal transducer 2 (TACSTD2), GA733 tumor associated antigen, and epithelial glycoprotein-1 (EGP-1), is a type I transmembrane protein highly expressed in carcinomas. Cell surface expression of TROP-2 is determined by flow cytometry using 488nm wavelength excitation and monitoring emitted fluorescence with a detector optimized to collect peak emissions at 565-605nm.
Applications:
Suitable for use in Flow Cytometry. Other applications not tested.
Recommended Dilutions:
Flow Cytometry: 10ul labels 1-2.5x10e5 cells. Validated for use with PC-3 cells.
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Cells may be Fc-blocked with 1ug of human IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction.
2. After blocking, add 10ul of T8663-95F to 1-2.5x10e5 cells and incubate for 30 minutes at RT.
3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (Catalog # F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human Lyse Buffer.
4. Resuspend the cells in Flow Cytometry Staining Buffer for analysis by flow cytometry. As a control for this analysis, cells in a separate tube should be treated with PE-labeled mouse IgG2A antibody.
Storage and Stability:
May be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C as an undiluted liquid. Stable for 12 months at 4 degrees C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.