"CCR4" Related Products

Expand

Human CCR4/Galpha15 Stable Cell Line-Chem-5

Cat.No.: CSC-RG0631
Cell Line Description: This human CCR4-expressing cell line is made in the Chem-5 host, which supports high levels of recombinant CCR4 expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between CCR4 and its ligands.
Background: CCR4 is a GPCR that is activated by chemokines TARC (thymus and activation regulated chemokine) and MDC (macrophage-derived chemokine). NK cells, Th2 cells, cutaneous memory T cells, macrophages and platelets express CCR4 and respond to TARC and MDC. CCR4-null mice display reduced airway hyperresponsiveness to fungal spores, lipopolysaccharide-induced endotoxic shock, and cardiac allograft rejection. In addition, antibody-mediated antagonism of MDC inhibits development of insulitis and diabetes in NOD mice.
Growth Properties: Adherent
Host Cell: Chem-5
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific silencing of CCR4. 2. Protein expression: CCR4 in this cell line has been validated by immunocytochemical staining.3. EC50 for calcium mobilization by TARC: 0.59 nM; Signal/noise at TARC Emax: 524; Z=0.85 with TARC at 2x EC50.
Application: Calcium flux assay, ligand binding assays
Sub-type: Chemokine
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 + 250μg/mL Hygromycin Plating medium: DMEM with 4.5 g/L glucose + 4mM glutamine + 10% heat-inactivated FBS + 1x NEAA + 10mM HEPES + 1x Pen-Strep Atmosphere: 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: 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: CCR4 chemokine (C-C motif) receptor 4 [ Homo sapiens ]
Official Symbol: CCR4
Synonyms: CCR4; chemokine (C-C motif) receptor 4; C-C chemokine receptor type 4; CC CKR 4; CD194; ChemR13; CKR4; CMKBR4; k5 5; CCR-4; C-C CKR-4; chemokine (C-C) receptor 4; K5-5; CC-CKR-4; HGCN:14099; MGC88293;
Gene ID: 1233
mRNA Refseq: NM_005508
Protein Refseq: NP_005499
MIM: 604836
UniProt ID: P51679
Chromosome Location: 3p24-p21.3
Pathway: Chemokine receptors bind chemokines, organism-specific biosystem; Chemokine signaling pathway, organism-specific biosystem; Chemokine signaling pathway, conserved biosystem; Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; Cytokine-cytokine receptor interaction, organism-specific biosystem; Cytokine-cytokine receptor interaction, conserved biosystem; G alpha (i) signalling events, organism-specific biosystem;
Function: C-C chemokine receptor activity; G-protein coupled receptor activity; chemokine receptor activity; receptor activity; signal transducer activity;

Online Inquiry

Note: There will be extra charge for optional service!
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

Please input "biomart" as verification code. Please review Creative BioMart's privacy policy for more information


Price does not include shipping and packaging costs.Request quote for final price.

Price Inquiry

Welcome! For price inquiries, please feel free to contact us through the form below. We will get back to you as soon as possible.