FlowCellect Chemokine Receptor CCR3 Surface Expression Identification and Quantification Kit

FlowCellect Chemokine Receptor CCR3 Surface Expression Identification and Quantification Kit

FlowCellect

100 Tests / Kit

Millipore’s FlowCellect™ GPCR Surface Identification kit is designed to provide an easier and more efficient way of characterizing GPCR expression on a given cell type of interest. This kit includes optimized buffers and a high affinity, specific antibody capable for detecting the GPCR target in question. Also included in the kit is a secondary antibody conjugate to R-Phycoerythrin, which is a bright fluorochrome ideal for measuring endogenous levels of GPCR expression on native cells. By following the steps outlined in the assay procedure, researchers can easily identify which native cell lines would be suitable for performing secondary assays in support of their structure-activity drug discovery efforts.

Additional test applications include the identification of high, medium, and low expressing cell cultures during the clonal selection process. When a researcher decides to build a stable cell line in-house, it is essential that the clone of choice will provide the GPCR surface expression needed to perform drug screening. Choosing the wrong clone can adversely affect a primary screen if the chosen clone cannot detect lower affinity compounds. This can lead to a lower hit rate, which can result in screening limitations when identifying potential blockbuster compounds.

Another added feature is the ability to quantitatively measure GPCR cell surface expression using flow cytometry as the method for detection. Provided in the kit are two vials of Millipore’s ChemiScreen™ GPCR cell lines, serving as assay controls. Using patented technology, Millipore’s ChemiScreen™ GPCR cell lines retain high surface expression as illustrated by the relatively high maximal number of binding sites (Bmax value) when compared to a native system.

By using Millipore’s FlowCellect™ Chemokine Receptor Surface Expression Identification kit, the researcher has the ability to identify a native GPCR expressing cell for biological assay development; the researcher can also easily select clonal stable cell lines for these GPCRs; finally, the researcher can monitor the changes of these receptor expression levels during a disease state.

The FlowCellect™ GPCR identification kits are designed for flow cytometry applications, which can be used for the identification and quantification of the surface expression level of GPCRs on any cells. The kit uses a GPCR antibody validated for flow cytometry to detect receptor expression, and also includes both positive and negative control cells with well characterized receptor expression levels for the purpose of quantification.

Chemokine receptors are very attractive targets because they are highly expressed on immune cells and regulate leukocyte activation and trafficking. Many of these targets have been indicated to play a key role in certain diseases states such as HIV infection, Rheumatoid Arthritis, and Diabetes. The expression levels of these receptors often reflect the different states of infectious diseases and inflammation.

Chemokine receptors are seven transmembrane G-Protein coupled receptors (GPCRs). The chemokine receptor family comprises of eighteen GPCR targets, in which ten are CC chemokine receptors, seven are CXC chemokine receptors, and one is a CX3C chemokine receptor. Currently, Millipore has launched 12 GPCR surface identification kits with an initial focus on the chemokine receptor family.

Eosinophils are major effector cells implicated in a number of chronic inflammatory diseases in humans, particularly bronchial asthma and allergic rhinitis. The chemokine receptor 3 (CCR3), a GPCR activated by chemokines eotaxin 1/2, MCP-3, MCP-4, and RANTES, mediates selective recruitment of eosinophils into tissue and thus has recently become an attractive biological target for therapeutic intervention [1]. It is widely expressed on cells involved in allergic inflammation, such as basophils, mast cells, airway epithelial cells, and potentially TH2 T-lymphocytes. Allergen-induced eosinophil infiltration into airways is reduced or eliminated in CCR3 and eotaxin 1/2 knockout mice and in mice treated with antibodies directed against CCR3 [2,3]. CCR3 antagonists are currently being developed for the treatment of asthma and other allergic disorders.

The control cells provided in the kit are based on the commercially available ChemiScreen™ GPCR stable cell lines from Millipore. Both the positive and negative control cell lines have been fully characterized using Pharmacology techniques such as radioligand binding assay to determine the receptor number for each cell type [4]. Using this information, now the researcher can determine the number of receptors per cell on any given cell type.

• Flow Cytometry
• in vitro Toxicology Assays

Human

• 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.
• Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

Lot to lot in-process testing on all kit components

• Signaling
• Chemokine Analysis

• 1. 100X Mouse anti-human CCR3 Monoclonal Antibody
• Clone ID [5E8]: - (Part No. CS202145) One 100uL vial
• 2. 500X Secondary Antibody conjugated to R-Phycoerythrin: - (Part No. CS202129) One 20uL vial
• 3. Antibody Diluent: - (Part No. CS202130) One 1mL vial
• 4. Fixation Buffer: - (Part No. CS202122) One 13mL bottle
• 5. 10X Wash Buffer: - (Part No. CS202123) One 13mL bottle
• 6. 5X Assay Buffer: - (Part No. CS202124) One 55mL bottle
• 7. Positive Control Cell Culture for CCR3 : - (Part No. CS202144) One vial supplied with 4x106 cells
• 8. Negative Control Cell Culture for CCR3 : - (Part No. CS202121) One vial supplied with 4x106 cells

4-8°C for antibodies and buffers; -80°C for positive and negative control cells; 6 Months

Mouse

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Reference Promo Code ZBIOS406 when placing your order*.

*This offer is valid until September 30, 2011. Guava Kits with catalogue numbers that begin with the number 4 are excluded from this promotion. Discount can not be combined with any other discount.

• Fluorescent
• ¹²⁵I

FlowCellect Chemokine Receptor CCR3 Surface Expression Identification and Quantification Kit

Flow Cellect

ExpressPro

Monoclonal Antibody

Research