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BioSafety Level	II
Organism	Human
SourceOrgan	Lung
Growth Properties	Adherent
Morphology	Epithelial
Description	A549 cells stably expressing CAS:9
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	Puromycin
Donor Age	58 year old
Donor Gender	Male
Freeze-Thaw Recovery	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 15mL sterile conical tube containing 5mL complete media. 5. Centrifuge cells at 1500rpm 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.
Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is PriGrow III medium available in ABM, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10%, 1 ug/ml puromycin (G264), and Penicillin/Streptomycin(G255). Atmosphere: air, 95%; Carbon dioxide (CO2), 5%. Temperature: 37.0°C.
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Disclaimer	1. The end user acknowledges that the Materials provided under abm’s Material Transfer Agreement (MTA) does not grant a license for commercial use or imply any ownership rights of any intellectual property rights relating to the Materials. 2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for functional freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 5. Please note that if the gene to be knockout may be essential to cell survival, abm will provide a hemizygous pool as the default deliverable to compensate for any lethal effects (unless otherwise requested).
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