"LPAR5" Related Products

Human LPAR5/Galpha15 Stable Cell Line-Chem-1

Cat.No.: CSC-RG0685
Cell Line Description: This human GPR92 -expressing cell line is made in the Chem-1 host, which supports high levels of recombinant GPR92 expression on the cell surface and contains high levels of the promiscuous G protein to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists at GPR92.
Background: Lysophosphatidic acid (LPA) is a lysophospholipid produced by activated platelets that inhibits adenylate cyclase and stimulates DNA synthesis, changes in cell morphology, and increases in intracellular calcium in a variety of cultured mammalian cells. A growing family of GPCRs mediates the biological effects of LPA. The first LPA receptors described, LPA1-3, share relatively high sequence similarity and are related to the sphingosine 1-phosphate receptors S1P1-6. Two more recently characterized LPA receptors, LPA4 and LPA6, are related to each other but are more distantly related to LPA1-3. LPA6, originally known as GPR92, mediates LPA-induced cytoskeletal changes, intracellular calcium flux and increased cAMP by coupling to G12/13, Gq, and Gs. In addition to binding LPA, LPA6 is also activated by several other lipid-derived molecules, the most potent of which is farnesyl pyrophosphate. GPR92 is expressed in several tissues, most prominently in the CD8+ intraepithelial lymphocytes of the gastrointestinal tract.
Growth Properties: Adherent
Host Cell: Chem-1
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific silencing of LPAR5.2. Protein expression: LPAR5 in this cell line has been validated by immunocytochemical staining.
Application: Calcium flux assay, ligand binding assays
Sub-type: Lysophospholipid
Propagation: Complete growth medium: DMEM with 4.5 g/L glucose + 4mM glutamine + 10% heat-inactivated FBS + 1x Nonessential amino acids + 10mM HEPES + 1x Pen-Strep + 250μg/mL Genetecin/G-418 Plating medium: DMEM with 4.5 g/L glucose + 4mM glutamine + 10% heat-inactivated FBS + 1x NEAA + 10mM HEPES + 1x Pen-StrepAtmosphere: air, 95%; carbon dioxide (CO2), 5%Temperature: 37 °C
Starting Cells From Frozen Cell Stock: 1. Immediately upon receipt, thaw cells or place cells in liquid nitrogen. Maintain frozen in liquid nitrogen for up to 5 years. 2. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37°C water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing growth media. Place the flask in a humidified incubator at 37°C with 5% CO2. 3. After 8-24 h, all live cells will be attached. Viability of the cells is expected to be 50-80%. At this time, replace media to remove residual DMSO, and return to incubator.
Subculturing: 1. When cells are approximately 80% confluent, passage the cells as follows: Remove media and wash once with HBSS without Ca2+ and Mg2+ (10 mL/T75). Add 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) and place in humidified incubator at 37°C with 5% CO2 until cells begin to round up and detach (5-10 minutes). Gently rap the side of the flask to dislodge the cells. Neutralize trypsin by addition of 4 mL Growth Media per 1 mL trypsin. 2. Cells are typically passaged 1:10 every 3-4 days. Passaging ratio may be varied according to requirements of the investigator.
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Freezing medium: DMEM with 4.5 g/L glucose + 4mM glutamine + 20% heat-inactivated FBS + 1x NEAA + 10mM HEPES + 1x Pen-Strep + 10% DMSO
Storage: Liquid nitrogen
Preservation: 1. Frozen stocks of cells should be prepared at the earliest passage possible after thawing, as follows: Count detached cells (prepared as in Subculture-Step 1). Centrifuge cells at 200 x g for 5 min. Resuspend cells at 5 x 10^6 cells/mL in Freezing Media (cell densities of 2-10 x 10^6 are also acceptable if necessary). Dispense 1 mL aliquots into cryopreservation vials. Freeze the cells by a controlled rate process, such as in an isopropanol-jacketed container placed at –70°C overnight. Store the vials in liquid nitrogen. 2. Use of cells immediately after thawing is feasible for some cell lines and is being further validated. Some cell lines may need to be passaged at least once after thawing prior to use in calcium flux assays. Cells should be resuspended in Plating Media for plating for calcium assay.
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: LPAR5 lysophosphatidic acid receptor 5 [ Homo sapiens ]
Official Symbol: LPAR5
Synonyms: LPAR5; lysophosphatidic acid receptor 5; G protein coupled receptor 92 , GPR92, GPR93; KPG_010; LPA5; LPA-5; LPA receptor 5; G protein-coupled receptor 92; G-protein coupled receptor 92; G-protein coupled receptor 93; GPR92; GPR93;
Gene ID: 57121
mRNA Refseq: NM_001142961
Protein Refseq: NP_001136433
MIM: 606926
UniProt ID: Q9H1C0
Chromosome Location: 12p13.31
Pathway: GPCRs, Class A Rhodopsin-like, organism-specific biosystem;
Function: G-protein coupled purinergic nucleotide receptor activity; G-protein coupled receptor activity; molecular_function; receptor activity; signal transducer activity;

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