ChemiSCREEN™ CCR9 Membrane Preparation

ChemiSCREEN™ CCR9 Membrane Preparation

• ChemiScreen
• Chemicon (Millipore)

1 mL

Full length Human CCR9

The CCR9 receptor is a GPCR expressed selectively in the thymus, lymph node, spleen, and the small bowel (SB). CCR9 binds to Thymus-expressed chemokine ligand (TECK / CCL25) (Zaballos et al. 1999; Youn et al., 1999; Yu et al., 2000), and is involved in directing the movement of mature T cells in the thymus (Carramolino et al., 2001b). CCR9 also plays an important role in the pathogenesis of SB Crohn's disease (Carramolino et al., 2001a; Papadakis et al, 2001). The CCR9 selective expression in the small bowel but not the colon indicates the role of CCR9 in the specialized immune response in the SB (Kunkel et al., 2000; Papadakis et al., 2000). It was also shown that CCR9-mediated antiapoptosis could help in the study of cell survival mechanism and T cell malfunctioning (Youn et al, 2002). Chemicon's CCR9 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of CCR9 interactions with its ligands. The membrane preparations exhibit an EC50 of 8 nM for TECK/CCL25 in a GTPgammaS binding assay.

GTPγS Binding and Radioligand Binding Assay.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

The protein encoded by this gene is a member of the beta chemokine receptor family. It is predicted to be a seven transmembrane protein similar to G protein-coupled receptors. Chemokines and their receptors are key regulators of the thymocytes migration and maturation in normal and inflammation conditions. The specific ligand of this receptor is CCL25. It has been found that this gene is differentially expressed by T lymphocytes of small intestine and colon, suggested a role in the thymocytes recruitment and development that may permit functional specialization of immune responses in different segment of the gastrointestinal tract. This gene is mapped to the chemokine receptor gene cluster region. Two alternatively spliced transcript variants have been described.

FUNCTION: SwissProt: P51686 # Receptor for chemokine SCYA25/TECK. Subsequently transduces a signal by increasing the intracellular calcium ions level. Alternative coreceptor with CD4 for HIV-1 infection.
SIZE: 369 amino acids; 42016 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Highly expressed in the thymus and low in lymph nodes and spleen.
SIMILARITY: SwissProt: P51686 ## Belongs to the G-protein coupled receptor 1 family.
MISCELLANEOUS: EC50 of SCYA25/TECK for isoform 1 is lower than for isoform 2.

Human

Chemicon

CCR9

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.Packaging method: Membrane protein was adjusted to 0.5 mg/ml in packaging buffer, rapidly frozen, and stored at -80°C.

peptide

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Chemokine

A

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• CCR9
• GPR28
• CC-CKR-9
• D6
• GPR-9-6
• CDw199
• CCR10
• CCR-9
• CMKBR9
• CCBP2

GPCR Membrane Preparations

ChemiSCREEN™ CCR9 Membrane Preparation

0.5 mg/mL

SPECIFICATIONS:
Kd for TECK/CCL25: ~8 nM in a GTPγS binding assay

Chem-1

ASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [³⁵S]-GTPγS (final concentration of 0.1 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl₂/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled TECK/CCL25 is added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30°C. The binding reaction is transferred to a GF/B filter plate (prewetted with water), and washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4. The plate is dried and counted.

One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific TECK-stimulated [³⁵S]-GTPγS binding.

The CCR9 membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.

Chem-1

Membranes

P51686

• NM_006641.2
• NM_031200.1

GPCR