"AVPR2" Related Products

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Human AVPR2-SNAP Stable Cell Line-HEK293

Cat.No.: CSC-RG0057
Cell Line Description: HEK293-HuAVPR2-SNAP cell line is a hypotriploid human cell line, which has been transfected with a Human arginine vasopressin receptor 2 (AVPR2) tagged in the N-terminus with SNAP to allow stably express of the human AVPR2 tagged in the N-terminus with SNAP protein. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: This gene encodes the vasopressin receptor, type 2, also known as the V2 receptor, which belongs to the seven-transmembrane-domain G protein-coupled receptor (GPCR) superfamily, and couples to Gs thus stimulating adenylate cyclase. The subfamily that includes the V2 receptor, the V1a and V1b vasopressin receptors, the oxytocin receptor, and isotocin and mesotocin receptors in non-mammals, is well conserved, though several members signal via other G proteins. All bind similar cyclic nonapeptide hormones. The V2 receptor is expressed in the kidney tubule, predominantly in the distal convoluted tubule and collecting ducts, where its primary property is to respond to the pituitary hormone arginine vasopressin (AVP) by stimulating mechanisms that concentrate the urine and maintain water homeostasis in the organism. When the function of this gene is lost, the disease Nephrogenic Diabetes Insipidus (NDI) results. The V2 receptor is also expressed outside the kidney although its tissue localization is uncertain. When these "extrarenal receptors" are stimulated by infusion of a V2 selective agonist (dDAVP), a variety of clotting factors are released into the bloodstream. The physiologic importance of this property is not known - its absence does not appear to be detrimental in NDI patients. The gene expression has also been described in fetal lung tissue and lung cancer associated with alternative splicing.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human AVPR2.2. Protein expression: AVPR2 expression in this cell line has been validated by WB.3. Functional validation: Ligand binding assay; Receptor dimerization assays; cAMP assay; Internalization assay
Sub-type: Vasopressin
Propagation: Complete growth medium: EMEM (EBSS), 2mM Glutamine, 1% Non Essential Amino Acids (NEAA), 10% FCS, 50U/ml penicillin, 50μg/ml streptomycin, 0.6mg/mL G418Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock: Thaw the vials of cryopreserved cells rapidly in a water bath at 37°C. Carefully transfer the cells to a cell culture vessel containing the appropriate volume of pre-warmed culture medium without G418 so that cell density reaches 4 x 10^4 to 8 x 10^4 cells/cm2. The day after, ensure that cells have properly attached and carefully replace with fresh culture medium supplemented with G418. Allow cells to grow until 80% confluency then transfer in a new cell culture vessel. Note that final DMSO concentration must not exceed 0.5%.
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.6. 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: AVPR2 arginine vasopressin receptor 2 [ Homo sapiens ]
Official Symbol: AVPR2
Synonyms: AVPR2; arginine vasopressin receptor 2; DIR, DIR3; vasopressin V2 receptor; nephrogenic diabetes insipidus; V2R; AVPR V2; antidiuretic hormone receptor; renal-type arginine vasopressin receptor; DI1; DIR; NDI; ADHR; DIR3; MGC126533; MGC138386;
Gene ID: 554
mRNA Refseq: NM_000054
Protein Refseq: NP_000045
MIM: 300538
UniProt ID: P30518
Chromosome Location: Xq28
Pathway: Aquaporin-mediated transport, organism-specific biosystem; Arf6 trafficking events, organism-specific biosystem; Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; G alpha (s) 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; 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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