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Multicolor flow cytometric analysis of CD192 (CCR2) expression on Mouse splenocytes. C57BL/6 (Top Plots) and BALB/c (Bottom Plots) mouse splenic leucocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse Ly-6G and Ly-6C antibody (Cat No. 562709) and with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; Left Plots) or Alexa Fluor™ 647 Rat Anti-Mouse CD192 (CCR2) antibody (Cat. No. 568347/568349; Right Plots) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of CD192 (CCR2) [or Ig Isotype control staining] versus Ly-6G and Ly-6C were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ Alexa Fluor™ 647 Rat Anti-Mouse CD192 (CCR2)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Y15-488.rMAb is a recombinant monoclonal antibody that specifically recognizes CD192 which is also known as C-C chemokine receptor type 2 (CCR2 or CC-CKR-2), Cmkbr2, or MCP-1 receptor (MCP-1-R). CD192 (CCR2) is a seven-transmembrane, G-protein-coupled, glycoprotein receptor that belongs to the beta chemokine receptor family. It is expressed on monocytes, macrophages, basophils and on some T cells and dendritic cells. Chemokines including CCL2 (Monocyte chemoattractant protein 1/MCP-1), CCL7 (MCP-3), or CCL12 (MCP-5) bind to and signal through CD192 (CCR2) to recruit monocytes and other leucocytes into inflammatory sites including tumors.
Development References (9)
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Engel DR, Maurer J, Tittel AP, et al. CCR2 mediates homeostatic and inflammatory release of Gr1(high) monocytes from the bone marrow, but is dispensable for bladder infiltration in bacterial urinary tract infection.. J Immunol. 2008; 181(8):5579-86. (Biology). View Reference
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Fujimura N, Xu B, Dalman J, Deng H, Aoyama K, Dalman RL. CCR2 inhibition sequesters multiple subsets of leukocytes in the bone marrow.. Sci Rep. 2015; 5:11664. (Biology). View Reference
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Geissmann F, Jung S, Littman DR. Blood monocytes consist of two principal subsets with distinct migratory properties.. Immunity. 2003; 19(1):71-82. (Biology). View Reference
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Griffith JW, Sokol CL, Luster AD. Chemokines and chemokine receptors: positioning cells for host defense and immunity.. Annu Rev Immunol. 2014; 32:659-702. (Biology). View Reference
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Kurihara T, Bravo R. Cloning and functional expression of mCCR2, a murine receptor for the C-C chemokines JE and FIC.. J Biol Chem. 1996; 271(20):11603-7. (Biology). View Reference
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Mack M, Cihak J, Simonis C, et al. Expression and characterization of the chemokine receptors CCR2 and CCR5 in mice.. J Immunol. 2001; 166(7):4697-704. (Biology). View Reference
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Sunderkötter C, Nikolic T, Dillon MJ, et al. Subpopulations of mouse blood monocytes differ in maturation stage and inflammatory response.. J Immunol. 2004; 172(7):4410-7. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.