CD326 (EpCAM) Monoclonal Antibody (1B7), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

Description: EpCAM (Epithelial cell adhesion molecule, CD326, KSA, TROP1) is a 40 kD cell-surface adhesion molecule participating in homophilic, calcium-independent cell-cell interactions. EpCAM is a type-I transmembrane protein, and is expressed primarily on the basolateral surface of most epithelia. Although normal epithelia express low levels of EpCAM, increased expression has been correlated with increased proliferation and progression to a mesenchymal phenotype. EpCAM has also been used as a diagnostic marker on circulating metastatic carcinoma cells, while cancer cells of non-epithelial origin do not express EpCAM. Applications Reported: This 1B7 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 1B7 antibody has been pre-titrated and tested by flow cytometric analysis of A549 human lung carcinoma cells. This can be used at 5 μL (0.06 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 μm post-manufacturing filtered. Ep-CAM is a ∽40 kD glycoprotein, which functions as an epithelial cell adhesion molecule. Ep-CAM mediates Ca2+-independent homotypic interactions. Formation of Ep-CAM-mediated adhesions has a negative regulatory effect on adhesions mediated by classic cadherins, which may have strong effects on the differentiation and growth of epithelial cells. Ep-CAM overexpression was suggested to be associated with enhanced epithelial proliferation. Ep-CAM is expressed in the majority of human carcinomas and is a marker for tumors of epithelial lineage.