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Human GPR39/Galpha15 Stable Cell Line-HEK293

Cat.No.: CSC-RG0716
Cell Line Description: This cloned human GPR39-expressing cell line is made in the HEK293 host, which supports high levels of recombinant GPR39 expression on the cell surface and supports optimal coupling of the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists at GPR39.
Background: GPR39 is a 7TM protein related to the G protein-coupled receptors for the gastrointestinal peptides ghrelin and motilin. Although GPCR39 has been reported to be a receptor for the peptide obestatin, GPR39 has been more thoroughly characterized as a receptor for Zn2+, as well as Cu2+ and Ni2+. Studies on GPR39-null mice implicate GPR39 in the control of insulin secretion, thereby indicating its potential as a target for treatment of diabetes.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific silencing of GPR39.2. Protein expression: GPR39 in this cell line has been validated by immunocytochemical staining.
Application: Calcium flux assay
Sub-type: Orphan
Propagation: Complete growth medium: DMEM/F12 with 2.5 mM glutamine, 10% heat-inactivated FBS, 1x Nonessential amino acids, 1x Pen-Strep, 250μg/mL Genetecin/G-418Plating medium: DMEM/F12 with 2.5 mM glutamine, 10% heat-inactivated FBS, 1x Nonessential amino acids, 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. 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 Accutase at 1 mL/T75 and keep at room temperature until cells begin to round up and detach (5-10 minutes). Gently rap the side of the flask to dislodge the cells. Neutralize Accutase by addition of 4 mL Growth Media per 1 mL Accutase. 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: 90% heat-inactivated FBS + 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: GPR39 G protein-coupled receptor 39 [ Homo sapiens ]
Official Symbol: GPR39
Synonyms: GPR39; MGC149541; G protein-coupled receptor 39; G-protein coupled receptor 39
Gene ID: 2863
mRNA Refseq: NM_001508.2
Protein Refseq: NP_001499.1
MIM: 602886
UniProt ID: O43194
Chromosome Location: 2q21-q22
Pathway: Class A/1 (Rhodopsin-like receptors); GPCR ligand binding; GPCRs, Class A Rhodopsin-like; Signal Transduction; Signaling by GPCR
Function: G-protein coupled receptor activity; metal ion binding

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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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