BD Horizon™ RY586 Rat Anti-Mouse CD192 (CCR2)
Clone Y15-488.rMAb (RUO)
Multicolor flow cytometric analysis of CD192 (CCR2) expression on viable Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 BD Horizon™ RY586 Rat IgG1, κ Isotype Control (Cat. No. 568822; Left Plots) or BD Horizon™ RY586 Rat Anti-Mouse CD192 (CCR2) antibody (Cat. No. 568535/568536; Right Plots) at 1 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 plot showing the correlated expression of CD192 (CCR2) [or Ig Isotype control staining] versus Ly-6G and Ly-6C was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 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 viable Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 BD Horizon™ RY586 Rat IgG1, κ Isotype Control (Cat. No. 568822; Left Plots) or BD Horizon™ RY586 Rat Anti-Mouse CD192 (CCR2) antibody (Cat. No. 568535/568536; Right Plots) at 1 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 plot showing the correlated expression of CD192 (CCR2) [or Ig Isotype control staining] versus Ly-6G and Ly-6C was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
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
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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 cells 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- 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
RY586 Rat IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
568822
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Cell Viability Solution RUO
Size
500 Tests
Cat No.
555815
568535 Rev. 1
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.
568535 Rev. 1
Format Details
RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
RY586
Yellow-Green 561 nm
564 nm
586 nm
568535 Rev.1
Citations & References
View product citations for antibody "568535" on CiteAb
Development References (9)
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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
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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
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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
568535 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.