HTS036M

ChemiSCREEN CCR9 Membrane Preparation

Human CCR9 GPCR membrane preparation for GTPγS Binding & Radioligand Binding Assays.

• 分類程式碼代碼: 41106514
• eCl@ss: 32161000
• NACRES: NA.84

屬性

• 生物源: human
• 品質等級: 100
• 重組細胞: expressed in Chem-1 cells
• 製造商／商標名: ChemiScreen; Chemicon^®
• 濃度: 0.5 mg/mL
• 技術: ligand binding assay: suitable (GTPγS); radioligand binding assay (RLBA): suitable
• NCBI登錄號: NM_006641.2; NM_031200.1
• UniProt登錄號: P51686
• 運輸包裝: dry ice

描述

一般說明

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.

應用

GTPγS Binding and Radioligand Binding Assay.

生化／生理作用

GPCR Class: A

Protein Target: CCR9

Target Sub-Family: Chemokine

品質

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

規格

Inucbation Conditions
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.

外觀

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.

儲存和穩定性

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

法律資訊

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

免責聲明

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.

安全資訊

• 儲存類別代碼: 12 - Non Combustible Liquids
• 水污染物質分類（WGK）: WGK 2