"CACNA1A" Related Products

Human CACNA1A/CACNB4/CACNA2D1 Stable Cell Line-CHO

Cat.No.: CSC-RI0071
Cell Line Description: CHO-HuCACNA1A/CACNB4/CACNA2D1 cell line is clonally-derived from a CHO cell line, which has been transfected with a human calcium channel, voltage-dependent, P/Q type, alpha 1A subunit (CACNA1A), a human calcium channel, voltage-dependent, beta 4 subunit (CACNB4) and a human calcium channel, voltage-dependent, alpha 2/delta subunit 1 (CACNA2D1) to allow stably express of the human CACNA1A, CACNB4 and CACNA2D1. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: The human CACNA1A gene encodes the pore-forming subunit of voltage-gated P/Q-type Ca2+ channels. Co-expression with the beta 4 and alpha2delta1 auxiliary subunits recapitulates function in heterologous expression (CHO cells). Cav2.1 channels, widely distributed in neurons, are therapeutic targets in treatment of inflammatory pain.
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific over-expression of human CACNA1A/CACNB4/CACNA2D1.2. Protein expression: CACNA1A/CACNB4/CACNA2D1 expression in this cell line has been validated by WB.3. Functional validation: manual patch clamp. mean Ca2+ current amplitude ± SEM at +10 mV= 0.42 ± 0.05 nA, n = 5 (14 additional cells tested had inward current amplitude<200>
Sub-type: Voltage-gated calcium channels
Propagation: Complete growth medium: Ham"s 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 1:4 to 1:8 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: CACNB4 calcium channel, voltage-dependent, beta 4 subunit [ Homo sapiens ]
Official Symbol: CACNB4
Synonyms: CACNB4; CAB4; CACNLB4; EA5; EIG9; EJM; EJM4; EJM6; calcium channel, voltage-dependent, beta 4 subunit; calcium channel voltage-dependent subunit beta 4; dihydropyridine-sensitive L-type, calcium channel beta-4 subunit; voltage-dependent L-type calcium channel subunit beta-4
Gene ID: 785
mRNA Refseq: NM_000726.3
Protein Refseq: NP_000717.2
MIM: 601949
UniProt ID: O00305
Chromosome Location: 2q22-q23
Pathway: Arrhythmogenic right ventricular cardiomyopathy (ARVC); Axon guidance; Cardiac muscle contraction; Depolarization of the Presynaptic Terminal Triggers the Opening of Calcium Channels; Developmental Biology; Dilated cardiomyopathy; Hypertrophic cardiomyopathy (HCM);
Function: contributes_to high voltage-gated calcium channel activity; protein binding; contributes_to voltage-gated calcium channel activity

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