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.
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.
Product Details
BD Pharmingen™
CCR2; Cmkbr2; MCP-1 receptor
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG1, κ
Mouse CD192 Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
12772
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
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
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Rat IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557731
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
BV421 Rat Anti-Mouse Ly-6G and Ly-6C RUO
Size
50 µg
Cat No.
562709
Cell Viability Solution RUO
Size
500 Tests
Cat No.
555815
568349 Rev. 2
Antibody Details
Y15-488.rMAb
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.
568349 Rev. 2
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
568349 Rev.2
Citations & References
View product citations for antibody "568349" on CiteAb
Development References (9)
-
Auffray C, Fogg D, Garfa M, et al. Monitoring of blood vessels and tissues by a population of monocytes with patrolling behavior.. Science. 2007; 317(5838):666-70. (Biology). View Reference
-
Bachelerie F, Ben-Baruch A, Burkhardt AM, et al. International Union of Basic and Clinical Pharmacology. [corrected]. LXXXIX. Update on the extended family of chemokine receptors and introducing a new nomenclature for atypical chemokine receptors.. Pharmacol Rev. 2014; 66(1):1-79. (Biology). View Reference
-
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
-
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
-
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
-
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
-
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
-
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
-
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
568349 Rev. 2
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.