"LPAR1" Related Products


Human LPAR1-FLAG Stable Cell Line-RH7777

Cat.No.: CSC-RG0283
Cell Line Description: RH7777-HuLPAR1-FLAG cell line is from Rattus sp. liver hepatoma buffalo, which has been transfected with a human lipid growth factor lysophosphatidic acid (LPAR1) tagged in the N-terminus with FLAG to allow stably express of the human LPAR1 tagged in the N-terminus with FLAG protein. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: The lipid growth factor lysophosphatidic acid (LPA) is responsible for cell signaling in diverse pathways including survival, proliferation, motility, and differentiation. LPA acts upon target cells by activating its cognate receptors, which belong to the G protein-coupled endothelial differentiation gene (EDG) family. Four mammalian cell surface LPA receptors have been identified so far: EDG-2 (LPA1), EDG-4 (LPA2), EDG-7 (LPA3) and LPA4 (GPR23/P2Y9). EDG-2 is the most widely expressed receptor, with high-level mRNAs in the colons, small intestine, placenta, brain and heart. Heterologous expression studies have shown that EDG-2 couples to both Gi/o and Gq to mediate PLC activation, inhibition of cAMP accumulation and activation of the MAPK pathway. EDG-2 deficient mice show phenotypic changes observed in psychiatric disease as well as impaired suckling behavior attributable to defective olfaction.
Growth Properties: Loosely adherent
Morphology: Epithelial
Host Cell: RH7777
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human LPA1.2. Protein expression: LPA1 expression in this cell line has been validated by WB.3. Functional validation: Functional assays.
Sub-type: Lysophospholipid
Propagation: Complete growth medium: DMEM, 10% FBS, 5 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 1:6 weekly is recommended Medium Renewal: 2 to 3 times per week
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 95%; DMSO, 5%
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: LPAR1 lysophosphatidic acid receptor 1 [ Homo sapiens ]
Official Symbol: LPAR1
Synonyms: LPAR1; lysophosphatidic acid receptor 1; EDG2, endothelial differentiation, lysophosphatidic acid G protein coupled receptor, 2; edg 2; Gpcr26; GPR26; LPA1; Mrec1.3; rec.1.3; vzg 1; LPA-1; LPA receptor 1; ventricular zone gene 1; lysophosphatidic acid receptor Edg-2; endothelial differentiation, lysophosphatidic acid G-protein-coupled receptor, 2; EDG2; VZG1; edg-2; vzg-1;
Gene ID: 1902
mRNA Refseq: NM_001401
Protein Refseq: NP_001392
MIM: 602282
UniProt ID: Q92633
Chromosome Location: 9q
Pathway: Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; G alpha (i) signalling events, organism-specific biosystem; G alpha (q) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; GPCR ligand binding, organism-specific biosystem; Gap junction, organism-specific biosystem; Gap junction, conserved biosystem;
Function: G-protein alpha-subunit binding; G-protein coupled receptor activity; PDZ domain binding; lysophosphatidic acid receptor activity; phospholipid binding; protein binding; receptor activity; signal transducer activity;

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