Flow cytometric analysis of CCR7 (CD197) on human peripheral blood leukocytes and lymphocytes. Whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD4 (clone RPA-T4, Cat. No. 564724), BD Horizon BV421 Mouse Anti-Human CD8 (clone RPA-T8, Cat. No. 562429 or 562428), FITC Mouse Anti-Human CD45RA (clone HI100, Cat. No. 561882 or 555488), and either APC Mouse IgG1, κ Isotype Control (Cat. No.340442) or APC Mouse Anti-Human CCR7 (CD197) (Cat. No. 566762 or 566763). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Flow cytometry and data analysis were performed using a BD FACS LSRFortessa™ X20 Flow Cytometer and FlowJo™ software. Plots 1 and 2: The contour plots showing Ig isotype control or CCR7 (CD197) staining versus side light-scatter were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Plot 3: The contour plot showing CCR7 (CD197) staining versus CD45RA was derived from CD4-positive T cell gated events with the forward and side light-scatter characteristics of viable lymphocytes. Please note that we have observed considerable donor-to-donor variation in the proportion of CCR7 (CD197)-negative CD4-positive lymphocytes. Plot 4: The contour plot showing CCR7 (CD197) staining versus CD45RA was derived from CD8-positive T cell gated events with the forward and side light-scatter characteristics of viable lymphocytes.
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Flow cytometric analysis of CCR7 (CD197) on human peripheral blood leukocytes and lymphocytes. Whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD4 (clone RPA-T4, Cat. No. 564724), BD Horizon BV421 Mouse Anti-Human CD8 (clone RPA-T8, Cat. No. 562429 or 562428), FITC Mouse Anti-Human CD45RA (clone HI100, Cat. No. 561882 or 555488), and either APC Mouse IgG1, κ Isotype Control (Cat. No.340442) or APC Mouse Anti-Human CCR7 (CD197) (Cat. No. 566762 or 566763). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Flow cytometry and data analysis were performed using a BD FACS LSRFortessa™ X20 Flow Cytometer and FlowJo™ software. Plots 1 and 2: The contour plots showing Ig isotype control or CCR7 (CD197) staining versus side light-scatter were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Plot 3: The contour plot showing CCR7 (CD197) staining versus CD45RA was derived from CD4-positive T cell gated events with the forward and side light-scatter characteristics of viable lymphocytes. Please note that we have observed considerable donor-to-donor variation in the proportion of CCR7 (CD197)-negative CD4-positive lymphocytes. Plot 4: The contour plot showing CCR7 (CD197) staining versus CD45RA was derived from CD8-positive T cell gated events with the forward and side light-scatter characteristics of viable lymphocytes.
Product Details
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)
5 µl
1236
AB_2869854
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 to APC under optimum conditions, and unconjugated antibody and free APC were removed.
Recommended Assay Procedures
Note: For best staining results, cell samples should be stained and maintained at 4-8°C before flow cytometric analysis.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- 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).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
BUV395 Mouse Anti-Human CD4 RUO
Size
100 Tests
Cat No.
564724
BV421 Mouse Anti-Human CD8 RUO
Size
25 Tests
Cat No.
562429
BV421 Mouse Anti-Human CD8 RUO
Size
100 Tests
Cat No.
562428
Antibody Details
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.
Format Details
APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC
Red 627-640 nm
651 nm
660 nm
Citations & References
Development References (8)
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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
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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
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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
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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
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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
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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
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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
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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
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.
