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Purified Mouse Anti-Human CCR7 (CD197)
Purified Mouse Anti-Human CCR7 (CD197)
Multicolor flow cytometric analysis of CCR7 (CD197) expression on human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and then stained with either purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; Left Plot) or purified Mouse Anti-Human CCR7 (CD197) antibody (Cat. No. 567209; Right Plot) at 0.25 µg/test. After washing, the cells were stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). After blocking with Normal Mouse Serum (ThermoFisher, Cat. No. 10410), the cells were further stained  with Alexa Fluor®647 Mouse Anti-Human CD4 (Cat. No. 557707) and BD Horizon™ BV421 Mouse Anti-Human CD45RA (Cat. No. 562885) antibodies. The two-color flow cytometric contour plot showing the correlated expression of CD45RA versus CCR7 (CD197) [or Ig Isotype control staining] was derived from CD4-positive T cell-gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD FACSCanto™ II Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Purified Mouse Anti-Human CCR7 (CD197)

Immunohistochemical analysis of CCR7 (CD197) expression by cells within human tonsil. Cold acetone-fixed frozen sections of human tonsil were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 550878; Left Panel) or Purified Mouse Anti-Human CCR7 (CD197) antibody (Cat. No. 567209; Right Panel) at 5 µg/ml for one hour at room temperature. A three-step staining procedure that employs a Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (Sav-HRP) (Cat. No. 550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissues were counterstained with hematoxylin. CCR7 (CD197) positive staining on tonsillar cells (Right Panel) is indicated by the intense brown labeling. Original magnification: 20X. Data shown on this Technical Data Sheet are not lot specific.

Multicolor flow cytometric analysis of CCR7 (CD197) expression on human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and then stained with either purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; Left Plot) or purified Mouse Anti-Human CCR7 (CD197) antibody (Cat. No. 567209; Right Plot) at 0.25 µg/test. After washing, the cells were stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). After blocking with Normal Mouse Serum (ThermoFisher, Cat. No. 10410), the cells were further stained  with Alexa Fluor®647 Mouse Anti-Human CD4 (Cat. No. 557707) and BD Horizon™ BV421 Mouse Anti-Human CD45RA (Cat. No. 562885) antibodies. The two-color flow cytometric contour plot showing the correlated expression of CD45RA versus CCR7 (CD197) [or Ig Isotype control staining] was derived from CD4-positive T cell-gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD FACSCanto™ II Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Immunohistochemical analysis of CCR7 (CD197) expression by cells within human tonsil. Cold acetone-fixed frozen sections of human tonsil were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 550878; Left Panel) or Purified Mouse Anti-Human CCR7 (CD197) antibody (Cat. No. 567209; Right Panel) at 5 µg/ml for one hour at room temperature. A three-step staining procedure that employs a Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (Sav-HRP) (Cat. No. 550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissues were counterstained with hematoxylin. CCR7 (CD197) positive staining on tonsillar cells (Right Panel) is indicated by the intense brown labeling. Original magnification: 20X. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
BLR2; CC chemokine receptor 7; CMKBR7; EBI1; EVI1; Epstein-Barr virus induced gene 1; MIP-3 beta receptor
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human CCR7 Transfected Cell Line
Flow cytometry (Routinely Tested), Immunohistochemistry-frozen (Tested During Development)
0.5 mg/ml
1236
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. Store undiluted at 4°C.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567209 Rev. 1
Antibody Details
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2-L1-A

The 2-L1-A monoclonal antibody specifically binds to the human CC chemokine receptor CCR7, also known as CD197, on the cell surface. CCR7 (previously known as BLR2, EBI1 and CMKBR7) is a seven-transmembrane, G-protein-coupled receptor specific for two CC chemokines: CCL19 (also known as MIP-3β, Exodus-3, and ELC) and CCL21 (also known as 6Ckine, Exodus-2 SLC, TCA4, and SCYA21). CCR7 mRNA is expressed mainly in lymphoid tissues including the spleen, lymph nodes and tonsil, in bone marrow, and on peripheral T and B lymphocytes, cord blood CD34-positive cells, and mature dendritic cells. In response to its cognate chemokines, CCR7 (CD197) mediates homing of leucocytes to secondary lymphoid tissues. Differential CCR7 (CD197) expression can be used to distinguish naive, central memory, and effector memory T cell subsets. The human CCR7 gene, unlike other CC chemokine receptor genes, has been mapped to chromosome 17 (region 17q12). Because the extracellular region of CCR2 (CD192) has significant sequence homology with CCR7 (CD197), BD Biosciences has confirmed that mAb 2-L1-A does not cross-react with CCR2 on the surface of transfected cells.

        

567209 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
567209 Rev.1
Citations & References
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Development References (8)

  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. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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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567209 Rev. 1

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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.