ALDH18A1 CRISPR Knockout 293T Cell Line
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Product Name
ALDH18A1 CRISPR Knockout 293T Cell Line -
Gene Name/Gene ID
ALDH18A1, 5832 -
Parental Cell Line
HEK293T -
Organism/Tissue/Cell Type
Human (H. sapiens), Kidney, CRISPR Knockout Cell Line -
Protocol
Culture MediaFor optimal cell culture, we recommend using PriCoat™ T25 Flasks (G299) or coating your preferred vessels with Applied Cell Extracellular Matrix (G422). Dulbecco's Modified Eagle Medium (DMEM), High Glucose (TM500) + 10% FBS (Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.
*Do not heat-inactivateThawing1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.
3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.
4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.
5. Incubate the cells at the recommended conditions.
SubcultureVolumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.
1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA (TM050) to the culture vessel.
2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.
3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.
6. Incubate the cells at the recommended conditions.
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QC
Presence of frameshift-inducing INDELs, confirmed by Sanger Sequencing
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Number of cells
1x106 cells / 1.0 ml -
Adherent/Suspension
Adherent, epithelial -
Biosafety level
II -
Storage Instructions
1. Visually examine the packaging containers for signs of leakage or breakage.
2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.
To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.
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Freezing conditions
- Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
- Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Please refer to our Cell Handling and Thawing Guidelines for detailed instructions on receiving, thawing, and culturing live cells:
https://www.abmgood.com/immortalized-cells-documents.html
Our warranty and return policy is outlined in abm’s Terms and Conditions, including details on product quality, limitations, and claims.
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
The number of times cells can divide depends on the cell type:
Please refer to the Growth Conditions section of your specific product page. This section will indicate whether Applied Cell Extracellular Matrix (G422), PriCoat™ flasks, or both are recommended for optimal cell attachment and growth, as requirements may vary by cell type.