HEK293 cells stably expressing RFP using CRISPR Knock In Technology at the Safe Harbor Site (AAVS1) Quantity: 10⁶ cells/1.0 mlOrganism: HumanSource Organ: KidneyBioSafety Level: IIGrowth Properties: AdherentMorphology: PolygonalSeeding Density: 30,000-40,000 cells/cm2, Recommended Split ratio of 1:2 to 1:4Markers: PuromycinPropagation: The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the completed growth medium, add the following components to the base medium with the final concentration: fetal bovine serum (TM999) to a final concentration of 10%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1% Carbon dioxide (CO2): 5%, Temperature: +37.0°C.Freeze Thaw: 1. Pre-warm complete media in a +37°C waterbath. 2. Remove the cryopreserved vial from the liquid nitrogen storage tank. 3. Thaw the cells quickly by placing the lower half of the vial into the +37°C water bath and remove after 60 seconds. There should still be a few ice crystals left after thawing. It is important not to over-thaw the cryovials as the presence of DMSO is toxic to the cells. 4. Re-suspend the cells in the vial and transfer the cell suspension into a 15 ml sterile conical tube containing 5 ml complete media. 5. Centrifuge cells at 200x g for 3 minutes to pellet. 6. Aspirate out the media, leaving cell pellet undisturbed. 7. Re-suspend pellet in fresh culture medium and plate in new culture vessel. 8. Incubate cultures at +37°C, 5% CO2. T3700 may show the presence of several floating cells after thawing. Within 48-72 hours these cells should revive. Quality Control: 1) Puromycin drug selection.2) RFP fluorescence expressionShipping Conditions: Dry IceStorage Conditions: -180°C