BB700 Rat Anti-Human CCR7 (CD197)

Consider BD Horizon RealBlue™ 705 (RB705) Reagents, a bright and clean fluorochrome alternative to BD Horizon™ BB700 and PerCP-Cy5.5 More Info

BD Horizon™ BB700 Rat Anti-Human CCR7 (CD197)

Name: BB700 Rat Anti-Human CCR7 (CD197)
Brand: BD Horizon™
SKU: 566437

Clone 3D12

(RUO)

Multicolor flow cytometric analysis of CD197 (CCR7) and CD45RA coexpression patterns on CD4+ and CD8+ human peripheral blood lymphocytes. Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD45RA (Cat. No. 562885), BD Horizon™ BB700 Rat Anti-Human CD197 (CCR7) (Cat. No. 566437/566438), FITC Mouse Anti-Human CD4 (Cat. No. 555346/561005/561842), and APC Mouse Anti-Human CD8 (Cat. No. 555369/561421/561952/561953) antibodies. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two-color flow cytometric dot plots showing the correlated expression of CD45RA versus CD197 (CCR7) were derived for CD4+ (Left Plot) or CD8+ (Right Plot) gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.

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Product Details

Brand: BD Horizon™

Alternative Name: CCR7, BLR-2, EBI-1, CMKBR7

Reactivity: Human (QC Testing)

Isotype: Rat IgG2a, κ

Immunogen: Human CCR7 protein

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Entrez Gene ID: 1236

RRID: AB_2744306

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™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.

Recommended Assay Procedures

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 or 566349).

When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent 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

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
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.
The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
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.
BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
Cy is a trademark of GE Healthcare.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

View Details

566437 Rev. 1

Antibody Details

3D12

The monoclonal antibody 3D12 reacts with the human CC chemokine receptor, CCR7. CCR7 (previously known as BLR-2, EBI-1 and CMKBR7), a seven-transmembrane, G-protein-coupled receptor, is the specific receptor for CC chemokines, MIP-3β/Exodus 3/ELC/ CCL19 and 6Ckine/Exodus 2/SLC/TCA4/CCL21. It has been shown that CCR7 mRNA is expressed mainly in lymphoid tissues including spleen, lymph nodes and tonsil. CCR7 mRNA was also detected in peripheral T and B lymphocytes, in bone marrow and cord blood CD34-positive cells and mature dendritic cells. The human CCR7 gene, unlike other CC chemokine receptor genes, has been mapped to chromosome 17q12. The immunogen used to generate 3D12 hybridoma was the N-terminus as well as parts of the second extracellular loop of human CCR7 protein. The monoclonal antibody 3D12 recognizes an epitope mapping to the N-terminus of human CCR7.

The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

566437 Rev. 1

Format Details

BB700

The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BB700

Excitation Source: Blue 488 nm

Excitation Max: 476 nm

Emission Max: 695 nm

566437 Rev.1

Citations & References

View product citations for antibody "566437" on CiteAb

Development References (15)

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Britschgi MR, Link A, Lissandrin TK, Luther SA. Dynamic modulation of CCR7 expression and function on naive T lymphocytes in vivo. J Immunol. 2008; 181(11):7681-7688. (Biology). View Reference
Burgstahler R, Kempkes B, Steube K, Lipp M. Expression of the chemokine receptor BLR2/EBI1 is specifically transactivated by Epstein-Barr virus nuclear antigen 2. Biochem Biophys Res Commun. 1995; 215(2):737-743. (Biology). View Reference
Forster R, Davalos-Misslitz AC, Rot A. CCR7 and its ligands: balancing immunity and tolerance. Nat Rev Immunol. 2008; 8(5):362-371. (Biology). View Reference
Kim CH, Pelus LM, White JR, Broxmeyer HE. Macrophage-inflammatory protein-3 beta/EBI1-ligand chemokine/CK beta-11, a CC chemokine, is a chemoattractant with a specificity for macrophage progenitors among myeloid progenitor cells. J Immunol. 1998; 161(5):2580-2585. (Biology). View Reference
Kurobe, H., Liu, et al. CCR7-dependent cortex-to-medulla migration of positively selected thymocytes is essential for establishing central tolerance. Immunity. 2006; 24(2):165-177. (Biology). View Reference
Lipp M, Burgstahler R, Muller G, et al. Functional organization of secondary lymphoid organs by the chemokine system. Curr Top Microbiol Immunol. 2000; 251:173-179. (Immunogen: Functional assay, Inhibition). View Reference
Ohl L, Mohaupt M, Czeloth N, et al. CCR7 governs skin dendritic cell migration under inflammatory and steady-state conditions. Immunity. 2004; 21(2):279-288. (Biology). View Reference
Ritter U, Wiede F, Mielenz D, Kiafard Z, Zwirner J, Korner H. Analysis of the CCR7 expression on murine bone marrow-derived and spleen dendritic cells.. J Leukoc Biol. 2004; 76(2):472-476. (Biology). View Reference
Sallusto F, Lenig D, Forster R, Lipp M, Lanzavecchia A. Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature. 1999; 401(6754):708-712. (Clone-specific: Flow cytometry). View Reference
Schweickart VL, Raport CJ, Godiska R, et al. Cloning of human and mouse EBI1, a lymphoid-specific G-protein-coupled receptor encoded on human chromosome 17q12-q21.2. Genomics. 1994; 23(3):643-650. (Biology). View Reference
Yanagihara S, Komura E, Nagafune J, Watarai H, Yamaguchi Y. EBI1/CCR7 is a new member of dendritic cell chemokine receptor that is up-regulated upon maturation. J Immunol. 1998; 161(6):3096-3102. (Biology). View Reference
Yoshida R, Imai T, Hieshima K, et al. Molecular cloning of a novel human CC chemokine EBI1-ligand chemokine that is a specific functional ligand for EBI1, CCR7. J Biol Chem. 1997; 272(21):13803-13809. (Biology). View Reference
Yoshida R, Nagira M, Imai T, et al. EBI1-ligand chemokine (ELC) attracts a broad spectrum of lymphocytes: activated T cells strongly up-regulate CCR7 and efficiently migrate toward ELC. Int Immunol. 1998; 10(7):901-910. (Biology). View Reference
Yoshida R, Nagira M, Kitaura M, Imagawa N, Imai T, Yoshie O. Secondary lymphoid-tissue chemokine is a functional ligand for the CC chemokine receptor CCR7. J Biol Chem. 1998; 273(12):7118-7122. (Biology). View Reference

View All (15)

566437 Rev. 1

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