The EpCAM Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. The particles contain a human EpCAM (NM_002354.3) driven by an EF1A promoter and a puromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the EpCAM Lentivirus
Two vials (500 µl x 2) of lentivirus at a titer ≥107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Formulation
The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
UniProt #
P16422
Background
Epithelial cell adhesion molecule (EpCAM, also known as CD90) is specific to epithelial cells and regulates cell proliferation and differentiation. It is used as an epithelial cell biomarker to detect circulating tumor cells. EpCAM belongs to the small GA733 protein family and is a homophilic transmembrane glycoprotein involved in cell-cell adhesion and tissue plasticity. It is a single-chain membrane-spanning protein with three major domains known as EpEX, involved in epidermal growth factor receptor (EGFR) mediated signaling pathways. Due to the versatile function of EpCAM and its crosstalk with other signaling pathways in cancer, EpCAM is considered an attractive target for cancer diagnosis, prognosis, and therapy.
Storage/Stability
Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.
Applications
Generation of a stable cell line expressing human EpCAM with puromycin selection
Shipping Temperature
-80°C
Notes
To generate a EpCAM stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve) for antibiotic selection of transduced cells. Visit: https://bpsbioscience.com/cell-line-faq for guidelines on performing a kill curve.
Biosafety The lentiviruses are produced with SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
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