Address: 45-1 Ramsey Road, Shirley, NY 11967, USA    USA: 1-631-559-9269  1-631-448-7888   Fax: 1-631-938-8127  Europe: 44-207-097-1828   Email: info@creative-biomart.com

Human KCNA4 Stable Cell Line-HEK293

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Chicken(1)
Human(3)
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His(3)
Cat.No.:
CSC-RI0129
Cell Line Description:
HEK293-HuKCNA4 cell line is a hypotriploid human cell line, which has been transfected with a human potassium voltage-gated channel, shaker-related subfamily, member 4 (KCNA4) to allow stably express of the human KCNA4. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background:
The human KCNA4 gene encodes the pore-forming subunit of Kv1.4, a voltage-gated potassium channel. Kv1.4 channels expressed in neurons are therapeutic targets in neuropathic pain.
Growth Properties:
Adherent
Morphology:
Epithelial
Host Cell:
HEK293
Cell Line Validation:
1. Gene expression: qPCR experiments determined specific expression of human KCNA4.2. Protein expression: KCNA4 expression in this cell line has been validated by WB.3. Functional validation: PatchXpress.
Application:
Therapeutic Area: pain & inflammation
Sub-type:
Voltage-gated potassium channels
Propagation:
Complete growth medium: DMEM/Complete growth medium: Ham's F12; 10% FBS; 500 μg/mL G418Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock:
1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37°C water bath for 2 minutes until nearly (80%) thawed. Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.2. Clean the outside of the vial with 70% ethanol before opening.3. Remove the cells from the vial and add slowly into a 15ml conical tube containing 10 ml pre-warmed media. 4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.6. Place the cells in the 37°C incubator with 5% CO2.7. Allow incubation for 3-4 days to reach confluence. The cells will re-attach to the surface over a period of several days in culture at 37°C.
Subculturing:
1. Remove and discard culture medium.2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.5. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended Medium Renewal: 2 to 3 times per week
Mycoplasma:
Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium:
Complete growth medium 90%; DMSO, 10%
Storage:
Liquid nitrogen
Preservation:
1. Detach cells from culture dish according to the Sub-Culture Procedure.2. Resuspend cells at a density of 5 x 10^6 cells/mL in freeze medium.Note: A T-75 culture flask typically yields enough cells for preparing two frozen vials.3. Aliquot 1 mL cells into cryogenic vials.4. Place vials in a freezing container and store at –80 °C overnight.5. Transfer vials to liquid nitrogen for long term storage. If properly stored, cells should remain stable for years.
Safety Considerations:
The following safety precautions should be observed.1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.2. No eating, drinking or smoking while handling the stable line.3. Wash hands after handling the stable line and before leaving the lab.4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.5. All waste should be considered hazardous.6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship:
Dry ice


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Optional requirements on this protein +Expand
C-fusion   N-fusion  Non-tagged
His   GST  Fc  Others
<1.0 eu/μg   <0.1 eu/μg  <0.01 eu/μg  Not required
Monomer Isolation   Dimer Isolation   Not required
>80% by SDS-PAGE   >90% by SDS-PAGE  >95% by SDS-PAGE  Others
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45-1 Ramsey Road, Shirley, NY 11967, USA
USA: 1-631-559-9269  1-631-448-7888
Europe: 44-207-097-1828
FAX: 1-631-938-8127
EMAIL: info@creative-biomart.com

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