"CACNA1H" Related Products

Human CACNA1H/CACNB2/CACNA2D1/KCNJ2 Stable Cell Line-HEK293

Cat.No.: CSC-RI0080
Cell Line Description: HEK293-HuCACNA1H/CACNB2/CACNA2D1/KCNJ2 cell line is a hypotriploid human cell line, which has been transfected with a human calcium channel, voltage-dependent, T type, alpha 1H subunit (CACNA1H), a human calcium channel, voltage-dependent, beta 2 subunit (CACNB2), a human calcium channel, voltage-dependent, alpha 2/delta subunit 1 (CACNA2D1) and a human potassium inwardly-rectifying channel, subfamily J, member 2 (KCNJ2) to allow stably express of the human CACNA1H, CACNB2, CACNA2D1 and KCNJ2. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: The human CACNA1H gene encodes the pore-forming subunit of Cav3.2, a voltage-gated T-type calcium channel. Coexpression with the beta 2 and alpha2delta1 auxiliary subunits recapitulates function in heterologous expression (HEK cells). Inclusion of the inwardly rectifying potassium channel allows the membrane potential to be altered with changes in extracellular K+.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific over-expression of human CACNA1H/CACNB2/CACNA2D1/KCNJ2.2. Protein expression: CACNA1H/CACNB2/CACNA2D1/KCNJ2 expression in this cell line has been validated by WB.3. Functional validation: FLIPR.
Sub-type: Inward rectifier potassium channels
Propagation: Complete growth medium: DMEM/F12/10% FBSAtmosphere: 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
Gene Name: KCNJ2 potassium inwardly-rectifying channel, subfamily J, member 2 [ Homo sapiens ]
Official Symbol: KCNJ2
Synonyms: KCNJ2; potassium inwardly-rectifying channel, subfamily J, member 2; inward rectifier potassium channel 2; IRK1; Kir2.1; IRK-1; hIRK1; inward rectifier K+ channel KIR2.1; cardiac inward rectifier potassium channel; LQT7; SQT3; ATFB9; HHIRK1; KIR2.1; HHBIRK1;
Gene ID: 3759
mRNA Refseq: NM_000891
Protein Refseq: NP_000882
MIM: 600681
UniProt ID: P63252
Chromosome Location: 17q24.3
Pathway: Activation of G protein gated Potassium channels, organism-specific biosystem; Activation of GABAB receptors, organism-specific biosystem; Cholinergic synapse, organism-specific biosystem; Classical Kir channels, organism-specific biosystem; G protein gated Potassium channels, organism-specific biosystem; GABA B receptor activation, organism-specific biosystem; GABA receptor activation, organism-specific biosystem;
Function: inward rectifier potassium channel activity; voltage-gated ion channel activity;

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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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