GFAT1 CRISPR Stable Knockout H1299 Cell Line

Cat. No.

T6199

Unit

1x10⁶ cells / 1.0 ml

Price

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Cat. No.	T6199
Name	GFAT1 CRISPR Stable Knockout H1299 Cell Line
Description	Glutamine-fructose-6-phosphate amidotransferase (GFAT1) is involved in the hexosamine biosynthesis pathway (HBP) which have been reported to have a critical role in the insulin resistance and progression of cancer. GFAT1 is the rate limiting step in the pathway thus is a target of interest for study. GFAT1 CRISPR Stable Knockout H1299 Cell Line was generated to study the effects of GFAT1. The cells may be rescued with treatment with exogenous GlcNAc (N-Acetylglucosamine) to maintain the propagation and glycosylation.
Organism	Human (H. sapiens)
Tissue	Lung
Donor History	Male, Caucasian, 43, Lymph node metastasis of the lung
Growth Properties	Adherent, epithelial
Unit	1x10⁶ cells / 1.0 ml
Storage Condition	Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions	Ship with dry ice.
Product Format	Frozen
Intended Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety	II
Certificate of Analysis	For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions	For optimal cell culture, we recommend using PriCoat™ T25 Flasks (G299) or coating your preferred vessels with Applied Cell Extracellular Matrix (G422) . RPMI 1640 Medium (TM503) + 10% FBS(Regular) + 10 mM N-Acetyl-D-glucosamine + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. Do not heat-inactivate
Unpacking and 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.
Thawing Protocol	1. 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.
Subculture Protocol	Volumes 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 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.
Cryopreservation	We recommend using serum-free CryoGuard™ Freezing Media (TM078) or, if serum is preferred, Cryopreservation Medium (TM024).
Population Doubling Time (h)	30
Warranty	abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period”.
Disclaimer	1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at licensing@abmgood.com. 2. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 3. We 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 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). 4. 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. 5. 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. 6. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Depositor	University of Southern California
Application	Research Use Only.
Material Citation	If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T6199

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	How should I handle live cells once I receive them?
	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 Following these guidelines will help ensure optimal cell viability and performance.

	What is your warranty or return policy?
	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.

How many times can cells divide?

The number of times cells can divide depends on the cell type:

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.

	Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
	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.

Darabedian, N., Gao, J., Chuh, K. N., Woo, C. M., & Pratt, M. R. (2018). The metabolic chemical reporter 6-azido-6-deoxy-glucose further reveals the substrate promiscuity of O-GlcNAc transferase and catalyzes the discovery of intracellular protein modification by O-glucose. Journal of the American Chemical Society, 140(23), 7092–7100. https://doi.org/10.1021/jacs.7b13488

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