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BV480 Rat Anti-Human CCR7 (CD197)
BV480 Rat Anti-Human CCR7 (CD197)
Multicolor flow cytometric analysis of CCR7 (CD197) and CD45RA coexpression patterns on CD4+ and CD8+ human peripheral blood lymphocytes. Human whole blood was stained with FITC Mouse Anti-Human CD45RA (Cat. No. 555488/561882), BD Horizon™ BV480 Rat Anti-Human CCR7 (CD197) (Cat. No. 566099/566170), PerCP-Cy™5.5 Mouse Anti-Human CD4 (Cat. No. 560650), and Alexa Fluor® 647 Mouse Anti-Human CD8 (Cat. No. 557708) 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 CCR7 (CD197) 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™ Cell Analyzer System.
Multicolor flow cytometric analysis of CCR7 (CD197) and CD45RA coexpression patterns on CD4+ and CD8+ human peripheral blood lymphocytes. Human whole blood was stained with FITC Mouse Anti-Human CD45RA (Cat. No. 555488/561882), BD Horizon™ BV480 Rat Anti-Human CCR7 (CD197) (Cat. No. 566099/566170), PerCP-Cy™5.5 Mouse Anti-Human CD4 (Cat. No. 560650), and Alexa Fluor® 647 Mouse Anti-Human CD8 (Cat. No. 557708) 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 CCR7 (CD197) 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™ Cell Analyzer System.
Product Details
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BD Horizon™
CCR7, BLR-2, EBI-1, CMKBR7
Human (QC Testing)
Rat IgG2a, κ
Human CCR7 protein
Flow cytometry (Routinely Tested)
5 µl
1236
AB_2739502
Aqueous buffered solution containing BSA and ≤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 with BD Horizon BV480 under optimum conditions, and unconjugated antibody and free BD Horizon BV480 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. 563794/566349).

For Immunofluorescence Applications:

The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480.  For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter.  

For epifluorescence microscopes with broad spectrum excitation sources,  the recommended excitation and emission filters are 445/20 nm and 485/20 nm bandpass filters, respectively.  For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Cy is a trademark of GE Healthcare.
  8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  10. 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.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Antibody Details
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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 BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

Format Details
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BV480
The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV480
Violet 405 nm
440 nm
479 nm
Citations & References
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View product citations for antibody "566099" on CiteAb

Development References (15)

  1. Birkenbach M, Josefsen K, Yalamanchili R, Lenoir G, Kieff E. Epstein-Barr virus-induced genes: first lymphocyte-specific G protein-coupled peptide receptors. Nature. 1993; 67(4):2209-2220. (Biology). View Reference
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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, Fluorescence activated cell sorting). View Reference
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.