"SCN10A" Related Products

Human SCN10A/SCN3B Stable Cell Line-CHO

Cat.No.: CSC-RI0173
Cell Line Description: CHO-HuSCN10A/SCN3B cell line is clonally-derived from a CHO cell line, which has been co-transfected with a human sodium channel, voltage-gated, type X, alpha subunit (SCN10A) and a human sodium channel, voltage-gated, type III, beta subunit (SCN3B) to allow stably express of the human SCN10A and SCN3B. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: The human SCN9A gene encodes the pore-forming subunit of Nav1.8, a voltage-gated sodium channel expressed in the peripheral nervous system including dorsal root ganglia. The SCN3B gene encodes an auxiliary beta -subunit that promotes surface expression. Nav1.8 channels are upregulated in chronic/inflammatory pain and are therapeutic targets in pain treatment.
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific expression of human SCN10A/SCN3B.2. Protein expression: SCN10A/SCN3B expression in this cell line has been validated by WB.3. Functional validation: FLIPR.
Application: Therapeutic Area: pain & inflammation
Sub-type: Voltage-gated sodium 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: SCN3B sodium channel, voltage-gated, type III, beta subunit [ Homo sapiens ]
Official Symbol: SCN3B
Synonyms: SCN3B; HSA243396; SCNB3; sodium channel, voltage-gated, type III, beta subunit; sodium channel subunit beta-3; voltage-gated sodium channel beta-3 subunit
Gene ID: 55800
mRNA Refseq: NM_001040151.1
Protein Refseq: NP_001035241.1
MIM: 608214
UniProt ID: Q9NY72
Chromosome Location: 11q23.3
Pathway: Axon guidance; Developmental Biology; Direct p53 effectors; Interaction between L1 and Ankyrins; L1CAM interactions
Function: ion channel binding; sodium channel regulator activity; voltage-gated sodium channel activity; voltage-gated sodium channel activity involved in regulation of cardiac muscle cell action potential

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