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PE Rat Anti-Human CCR7 (CD197)
PE Rat Anti-Human CCR7 (CD197)

Flow cytometric analysis of CCR7 (CD197) expression on CD4 and CD8-positive human peripheral lymphocytes. Human PBMC were stained with 20 ul/test of PE Rat Anti-Human CCR7 (CD197) (Cat. No.561008/552176) and FITC Mouse Anti-Human CD45RA (Cat. No. 555488). Bivariate dot plots are derived from the CD4-postive (based on staining with APC Mouse Anti-Human CD4, Cat. No. 555349, left panel) and CD8-positive (based on staining with APC Mouse Anti-Human CD8, Cat. No. 555369, right panel) lymphocyte gated populations.

Flow cytometric analysis of CCR7 (CD197) expression on CD4 and CD8-positive human peripheral lymphocytes. Human PBMC were stained with 20 ul/test of PE Rat Anti-Human CCR7 (CD197) (Cat. No.561008/552176) and FITC Mouse Anti-Human CD45RA (Cat. No. 555488). Bivariate dot plots are derived from the CD4-postive (based on staining with APC Mouse Anti-Human CD4, Cat. No. 555349, left panel) and CD8-positive (based on staining with APC Mouse Anti-Human CD8, Cat. No. 555369, right panel) lymphocyte gated populations.

Product Details
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BD Pharmingen™
CCR7, BLR-2, EBI-1, CMKBR7
Human (QC Testing)
Rat IgG2a, κ
Human CCR7 protein
Flow cytometry (Routinely Tested)
20 µl
1236
AB_2033950
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

The PE-conjugated 3D12 antibody is convenient for the immunofluorescent staining and flow cytometric analyses of human leukocytes and cell lines that express CCR7 (see figure). Chemokine receptors are known to internalize during manipulation resulting in low frequency expression. Immunophenotyping studies of chemokine receptors need to be performed on freshly collected whole blood (<24 hours). Incubation with the antibody should be done at 4°C in the dark. Cellular manipulation, such as Ficoll separation, freezing, or exposure to cold temperatures prior to staining have been shown to cause a decrease in staining intensity and inconsistent results.

A multiple-step staining procedure is strongly recommended to amplify immunofluorescent signals in the flow cytometric analysis of human CCR7 expression. Purified anti-human CCR7, clone 3D12, Cat. No. 552175 and a 3-step staining procedure is recommended for detecting low frequency expression.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561008 Rev. 2
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.

561008 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
561008 Rev.2
Citations & References
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Development References (9)

  1. 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
  2. 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
  3. 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
  4. 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). View Reference
  5. 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
  6. 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
  7. 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). View Reference
  8. 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
  9. 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
View All (9) View Less
561008 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.