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Immortalized Mouse Epidermal Keratinocytes (CDC25A F7)

SKU : BHC10900768

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Applied Biological Materials (abm) Inc.

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Mfr.No.	T0299
Description	Cell division cycle 25 (CDC25) family of phosphatases consists of CDC25A, CDC25B and CDC25C in which regulates the cell cycle progression, and thus are important for the activation of cell cycle checkpoints in response to DNA damage. CDC25A degradation has been associated with cell cycle arrest. Immortalized Mouse Epidermal Keratinocytes (CDC25A F7) Cells were isolated from the Cdc25afl/fl/Cre-?ice. The cells are suitable as a control for the mutant cells, in which CDC25A is not expressed, Immortalized Mouse Mutant Epidermal Keratinocytes (CDC25A 8b) Cells (Cat. No. T0298).
Cell Type	Immortalized Cells
Biosafety Level	2
Depositor	Creighton University
Disclaimer	1. All test parameters provided in the CoA are conducted using abm's standardized culture system and 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 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. 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. 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 or any other non-RUO application(s). 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 5. 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".
Donor History	C57/BL6 Cdc25afl/fl/Cre- mouse
Expression Level	Arnt2 and Mash1 (Immunofluorescence); Nestin, Sox2, and Pax6 (RT-PCR)
Growth Properties	Adherent, cobble-stone
Growth Conditions	Use of PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Thaw the cells with PriGrow XI (TM011) + 8% FBS + 60 ? CaCl? (added fresh each time) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 8% CO? for optimal attachment. After 24h, change media to PriGrow XI (TM011) + 8% chelexed FBS + 1.4% FBS + 5 ng/ml rhFGF7 (Z100595) (freshly added) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 8% CO? Steps to thaw the cells: 1. Thaw the cells in RT water until only a few ice crystals are left 2. Put the cells in 5 ml of RT PriGrow XI (TM011) + 8% FBS 3. Centrifuge the cells 1500 rpm for 5 min (need this step to remove inhibitors for attachment) 4. Resuspend the pellet in RT Prigrow XI (TM011) + 8% FBS + 1% P/S with freshly added 60 ? CaCl? and put the cells in a culture vessel at 37.0?, 8% CO? 5. After 18h-24h, change to PriGrow XI (TM011) + 8% chelexed FBS + 1.4% FBS + 5 ng/ml rhFGF7 (Z100595) (freshly added) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 8% CO? for growth of the cells.
Quantity	1x106 cells / 1.0 ml
Tissue	Skin
Notes	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?, 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? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism	Mouse (M. musculus)
Population Doubling	24 hours
Species	Mouse (M. musculus)
Propagation	PriGrow X Series Medium (TM0861) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO?. Thaw in T25, subculture with Accutase:PBS (ratio 1:1) for 3 minutes.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.

Applications Range	Research Use Only.

Shipping	Ship with dry ice.
Product Format	Frozen
Product 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.
Storage	Vapor phase of liquid nitrogen, or below -130?.

Thawing Protocol	1. Thaw cells quickly in a 37? 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?, 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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SKU	Supplier No.	Size	Your price	Quantity
BHC10900768	T0299	1x10^6 cells / 1.0 ml	-		Add to Quote

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