Staining of C57BL/6 splenocytes at 37°C with Anti-Mouse CD3e FITC (cat. 11-0031) and 1.0 µg of Anti-Mouse CD197 (CCR7) eFluor® 450. Cells in the lymphocyte gate were used for analysis.
Contents: Anti-Mouse CD197 (CCR7) eFluor® 450 (Pacific Blue® replacement) Catalog Number: 48-1971 Concentration: 0.2 mg/ml Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material. Clone: 4B12 Host/Isotype: Rat IgG2a, κ
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Size
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48-1971-80
25 ug
48-1971-82
100 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
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*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
Please see our website for continuous updates regarding the use of this product.
The 4B12 monoclonal antibody reacts with mouse CCR7, also known as EBI-1 and CD197. For optimal visualization of CCR7 expression on different cell types it is necessary to use multi-color staining to discriminate different cell subsets (please see Technical Data Sheets for recommended concentration for individual formats). To address specificity, the staining profile of 4B12 has been compared to a polyclonal antibody generated against a CCR7 peptide (Bjorkdahl et al). This analysis confirms that the polyclonal antibody and 4B12 stain similar populations of cells. Furthermore, 4B12 stains mouse CCR7-GFP fusion protein-transfected RBL cells (see data in cat. 14-1971).
CCR7 is a chemokine receptor for the chemokines CCL19 (CKß11, ELC, MIP3ß, Scya19, Exodus-3) and CCL21 (CKß9, SLC, MIP2ß, Scya21, Exodus-2). In recent years, the role of chemokines in directing the migration of lymphocytes has been well-characterized. One of the most important mediators of homeostatic trafficking of naïve T cells to secondary lymphoid organs (SLO) is the chemokine receptor CCR7. Binding of its ligands, CCL19 and CCL21, mediates the transendothelial migration of T cells across high endothelial venules into SLO. It has also been demonstrated that CCR7 plays a role in the localization of dendritic cells and B cells during an immune response.
In addition to its significant role in the chemotaxis of lymphocytes, human CCR7 has also been recognized as a marker for a distinct subset of memory T cells, the central memory (TCM) population. These cells are characterized by the expression of CCR7 and CD62L and reside within peripheral lymphoid organs.
CCR7 also plays a role in thymocyte development and its deficiency leads to disturbed thymic architecture, aberrant T cell development, and limited thymocyte expansion.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This 4B12 antibody has been reported for use in flow cytometric analysis.
Applications Tested
This 4B12 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 1 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
eFluorTM 450 is a replacement for Pacific Blue®. eFluorTM 450 emits at 456 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome.
References
Ritter, U, Wiede, F, Mielenz, D, Kiafard, Z, Zwirner, J, and Korner H. 2004. Analysis of the CCR7 expression on murine bone arrow-derived and spleen dendritic cells. Journal of Leukocyte Biology. 76: 472-476. (4B12, FC, Pubmed)
Bjorkdahl O, Barber KA, Brett SJ, Daly MG, Plumpton C, Elshourbagy NA, Tite JP, Thomsen LL. 2003. Characterization of CC-chemokine receptor 7 expression on murine T cells in lymphoid tissues. Immunology. 110(2):170-9.
Waldner H, Sobel RA, et al. 2006. The autoimmune diabetes locus Idd9 regulates development of type 1 diabetes by affecting the homing of islet-specific T cells. J Immunol. 176(9):5455-62. (4B12, FC, PubMed)
Ohl L, Mohaupt M, Czeloth N, Hintzen G, Kiafard Z, Zwirner J, Blankenstein T, Henning G, Forster R. 2004. CCR7 governs skin dendritic cell migration under inflammatory and steady-state conditions.
Immunity. 21(2):279-88. (4B12, FC, Pubmed)
