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Human VIPR1-SNAP Stable Cell Line-CHO

Cat.No.: CSC-RG0055
Cell Line Description: CHO-HuVIPR1-SNAP cell line is clonally-derived from a CHO cell line, which has been transfected with a Human vasoactive intestinal peptide receptor 1 (VIPR1) tagged in the N-terminus with SNAP to allow stably express of the human VIPR1 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 a receptor for vasoactive intestinal peptide, a small neuropeptide. Vasoactive intestinal peptide is involved in smooth muscle relaxation, exocrine and endocrine secretion, and water and ion flux in lung and intestinal epithelia. Its actions are effected through integral membrane receptors associated with a guanine nucleotide binding protein which activates adenylate cyclase. Several transcript variants encoding different isoforms have been found for this gene.
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human VIPR1.2. Protein expression: VIPR1 expression in this cell line has been validated by WB.3. Functional validation: Ligand binding assay; Receptor dimerization assays; cAMP assay; Phospho-ERK assay; Internalization assay
Sub-type: Vasoactive Intestinal Peptide
Propagation: Complete growth medium: Ham"s F12, 10% dFBS, 50U/ml penicillin, 50μg/ml streptomycin, 2mM Hepes Atmosphere: 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: 45% Medium; 45% FCS; 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: VIPR1 vasoactive intestinal peptide receptor 1 [ Homo sapiens ]
Official Symbol: VIPR1
Synonyms: VIPR1; vasoactive intestinal peptide receptor 1; vasoactive intestinal polypeptide receptor 1; HVR1; RDC1; VIP and PACAP receptor 1; VPAC1; VPAC1R; VIP receptor, type I; PACAP type II receptor; pituitary adenylate cyclase activating polypeptide receptor, type II; II; V1RG; VIPR; VIRG; VAPC1; VIP-R-1; VPCAP1R; PACAP-R2; PACAP-R-2; FLJ41949;
Gene ID: 7433
mRNA Refseq: NM_001251882
Protein Refseq: NP_001238811
MIM: 192321
UniProt ID: P32241
Chromosome Location: 3p22
Pathway: Class B/2 (Secretin family 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 B Secretin-like, organism-specific biosystem; Glucagon-type ligand receptors, organism-specific biosystem; Glucocorticoid receptor regulatory network, organism-specific biosystem;
Function: receptor activity; signal transducer activity; vasoactive intestinal polypeptide receptor activity;

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