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Human AVPR1A-FLAG Stable Cell Line-CHO

Cat.No.: CSC-RG0414
Cell Line Description: CHO-HuAVPR1A-FLAG is clonally-derived from a CHO-K1 cell line which has been transfected with a human arginine vasopressin receptor 1A (AVPR1A) tagged in the N-terminus with FLAG to allow stably express of the human AVPR1A tagged in the N-terminus with FLAG. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: V1A, a G protein coupled receptor, also called arginine vasopressin 1A (AVPR1A), is a receptor for neurohypophyseal peptide [Arg8]-vasopressin. V1A has shown to be highly expressed in the posterior pituitary gland, where it plays an essential role in controlling the water content of the body by acting on the kidney to increase water and sodium absorption. It is also abundant in the periphery in vascular smooth muscle, myometrium and the bladder, where it also acts to mediate contraction. Other functions include glycogen breakdown in the liver, inducing platelet activation, and triggering the release of corticotrophin from the anterior pituitary gland. Vasopressin is used clinically to treat diabetes insipidus. V1a receptor signals through phosphatidylinositol hydrolysis to mobilize intracellular Ca2+. Studies have shown V1A receptor antagonists to selectively block aggressive behavior in hamsters. Repeat variations in V1A has also been associated with the risk of autism, level of musical cognition, and level of altruism.
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO-K1
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human AVPR1A.2. Protein expression: AVPR1A expression in this cell line has been validated by WB.3. Functional validation: Functional assays.
Sub-type: Vasopressin
Propagation: Complete growth medium: DMEM/F12, 10% FBS, 10 μg/mL puromycinAtmosphere: 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: AVPR1A arginine vasopressin receptor 1A [ Homo sapiens ]
Official Symbol: AVPR1A
Synonyms: AVPR1A; arginine vasopressin receptor 1A; AVPR1; vasopressin V1a receptor; V1aR; AVPR V1a; V1a vasopressin receptor; antidiuretic hormone receptor 1A; SCCL vasopressin subtype 1a receptor; V1-vascular vasopressin receptor AVPR1A; vascular/hepatic-type arginine vasopressin receptor;
Gene ID: 552
mRNA Refseq: NM_000706
Protein Refseq: NP_000697
MIM: 600821
UniProt ID: P37288
Chromosome Location: 12q14-q15
Pathway: Calcium signaling pathway, organism-specific biosystem; Calcium signaling pathway, conserved biosystem; Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; G alpha (q) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; GPCR ligand binding, organism-specific biosystem; GPCRs, Class A Rhodopsin-like, organism-specific biosystem;
Function: G-protein coupled receptor activity; V1A vasopressin receptor binding; peptide hormone binding; protein binding; protein kinase C binding; receptor activity; signal transducer activity; vasopressin receptor activity;

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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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