HeLa-EGFRvIII
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Description
HeLa-EGFRvIII cells were generated from the human cervical cancer cell line HeLa by transfection with an expression vector encoding EGFRvIII with a C-terminal FLAG tag, followed by selection in medium containing geneticin. Expression of EGFRvIII was confirmed by flow cytometry using cells that were fixed and permeabilized (Figure 1).
Product Overview
Composition
Suspension of cells in 0.5 ml - 1 ml of 90% FBS + 10% DMSO.
Storage
Store vial in liquid nitrogen immediately upon receipt.
Thawing
Partially immerse the vial in a 37°C water bath with gentle shaking until most of the medium is thawed. In a tissue culture hood, add 1 ml of pre-warmed culture medium into the vial and immediately transfer the contents of the vial to a centrifuge tube containing 5-10 ml of pre-warmed culture medium. Centrifuge the tube at room temperature for 5 minutes, aspirate the supernatant and suspend the cell pellet in 5-10 ml of pre-warmed culture medium.
Culture
HeLa-EGFRvIII is an adherent cell line. Culture the cells in DMEM containing 10% FBS and 1 mg/ml geneticin using a humidified incubator set to 5% CO2. When the cell monolayer is nearly confluent, use trypsin-EDTA to detach the cells from the culture flask and immediately neutralize the trypsin by adding fresh, pre-warmed culture medium to the cells. Cells should be passaged at least twice per week.
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Figure 1
PM-EGFRvIII-HELA cells were fixed, permeabilized and stained with an anti-FLAG antibody or an isotype control antibody. The FLAG antibody bound to over 95% of the PM-EGFRvIII-HELA cells.
For Research Use Only. Not for use in diagnostic procedures.