BB515 Rat Anti-Mouse CD195

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BD Horizon™ BB515 Rat Anti-Mouse CD195

Name: BB515 Rat Anti-Mouse CD195
Brand: BD Horizon™
SKU: 565093

Clone C34-3448

(RUO)

Two-color flow cytometric analysis of CD195 expression on mouse peritoneal exudate cells. Mouse peritoneal exudate cells (PEC) were isolated 3 days post-stimulation by intraperitoneal injection of a 3% thioglycollate solution. The PEC were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD11b (Cat. No. 553312/561690) and either BD Horizon™ BB515 Rat IgG2c, κ Isotype Control (Cat. No. 565091; Left Panel) or BD Horizon BB515 Rat Anti-Mouse CD195 (Cat. No. 565093; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD195 (or Ig Isotype control staining) versus CD11b were derived from gated events with the forward and side light-scatter characteristics of viable PEC. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

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Product Details

Brand: BD Horizon™

Alternative Name: Ccr5; chemokine (C-C) receptor 5; C-C CKR-5; CC-CKR-5; CCR-5; AM4-7

Reactivity: Mouse (QC Testing)

Isotype: Rat IgG2c, κ

Immunogen: Mouse CCR5 aa. 9-30

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

RRID: AB_2739063

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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 with BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

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).
BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

BB515 Rat IgG2c, κ Isotype Control RUO

Size 50 µg Cat No. 565091

APC Rat Anti-CD11b RUO

Size 0.1 mg Cat No. 553312

View Details

565093 Rev. 2

Antibody Details

C34-3448

The C34-3448 monoclonal antibody specifically binds to CD195 which is also known as, C-C chemokine receptor type 5 (CCR5). CD195 is a seven transmembrane-spanning G-protein-coupled receptor that belongs to the β-chemokine receptor family. CD195 regulates lymphocyte chemotaxis activation and transendothelial migration during inflammation. It signals in response to at least three chemokines: CCL3/MIP-1α, CCL4/MIP-1β, and CCL5/RANTES. CD195 is expressed on macrophages and some T-lymphocytes.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

565093 Rev. 2

Format Details

BB515

The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BB515

Excitation Source: Blue 488 nm

Excitation Max: 490 nm

Emission Max: 515 nm

565093 Rev.2

Citations & References

Development References (10)

Boring L, Gosling J, Monteclaro FS, Lusis AJ, Tsou CL, Charo IF. Molecular cloning and functional expression of murine JE (monocyte chemoattractant protein 1) and murine macrophage inflammatory protein 1alpha receptors: evidence for two closely linked C-C chemokine receptors on chromosome 9. J Biol Chem. 1996; 271(13):7551-7558. (Biology). View Reference
Hayakawa Y, Smyth MJ. CD27 dissects mature NK cells into two subsets with distinct responsiveness and migratory capacity. J Immunol. 2006; 176(3):1517-1524. (Clone-specific: Flow cytometry). View Reference
Kim SH, Cleary MM, Fox HS, Chantry D, Sarvetnick N. CCR4-bearing T cells participate in autoimmune diabetes. J Clin Invest. 2002; 110(11):1675-1686. (Clone-specific: Flow cytometry). View Reference
McGuirk P, McCann C, Mills KH. Pathogen-specific T regulatory 1 cells induced in the respiratory tract by a bacterial molecule that stimulates interleukin 10 production by dendritic cells: a novel strategy for evasion of protective T helper type 1 responses by Bordetella pertussis. J Exp Med. 2002; 195(2):221-231. (Clone-specific: Flow cytometry). View Reference
McLoughlin RM, Jenkins BJ, Grail D, et al. IL-6 trans-signaling via STAT3 directs T cell infiltration in acute inflammation. Proc Natl Acad Sci U S A. 2005; 102(27):9589-9594. (Clone-specific: Flow cytometry). View Reference
Meyer A, Coyle AJ, Proudfoot AE, Wells TN, Power CA. Cloning and characterization of a novel murine macrophage inflammatory protein-1 alpha receptor. J Biol Chem. 1996; 271(24):14445-14451. (Biology). View Reference
Nakae S, Iwakura Y, Suto H, Galli SJ. Phenotypic differences between Th1 and Th17 cells and negative regulation of Th1 cell differentiation by IL-17. J Leukoc Biol. 2007; 81(5):1258-1268. (Clone-specific: Flow cytometry). View Reference
Napolitano M, Seamon KB, Leonard WJ. Identification of cell surface receptors for the Act-2 cytokine. J Exp Med. 1990; 172(1):285-289. (Biology). View Reference
Zozulya AL, Reinke E, Baiu DC, Karman J, Sandor M, Fabry Z. Dendritic cell transmigration through brain microvessel endothelium is regulated by MIP-1alpha chemokine and matrix metalloproteinases. J Immunol. 2007; 178(1):520-529. (Clone-specific: Flow cytometry). View Reference
van Deventer HW, Wu QP, Bergstralh DT, et al. C-C chemokine receptor 5 on pulmonary fibrocytes facilitates migration and promotes metastasis via matrix metalloproteinase 9. Am J Pathol. 2008; 173(1):253-264. (Clone-specific: Flow cytometry). View Reference

View All (10)

565093 Rev. 2

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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.