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Description: The 3D12 monoclonal antibody reacts with human CCR7, also known as EBI-1 and CD197. CCR7 is a member of the G-protein-coupled chemokine receptor family with seven membrane-spanning domains and functions as a receptor for 6Ckine/SLC (secondary lymphoid-tissue chemokine), CCL19 and CCL21. CCR7 has been shown to be internalized via clathrin-coated pits and the majority recycled back to the plasma membrane. CCR7 is expressed on T cells and can be used to distinguish populations of naive from central and effector memory T cells. CCR7 has been shown to play a role in migration of memory T cells to inflamed tissue. Expression of CCR7 is also found on DC's. During DC maturation CCR7 expression increases and is thought to be involved in a variety of functions: chemotaxis to the lymph node, cellular architecture, rate of endocytosis, survival and maturation. Expression of CCR7 on the cell surface can be down regulated upon ligand binding.
Applications Reported: This 3D12 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This 3D12 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.5 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 10^5 to 10^8 cells/test.
It is recommended that the staining incubation time be increased to at least 45 minutes at 2-8°C.
Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
CCR7 is a member of the G protein coupled receptor family, which is a subfamily of chemokines. CCR7 was identified to be induced by the Epstein Barr virus (EBV), and is thought to be a mediator of EBV effects on B lymphocytes. CCR7 has been reported to be expressed in blood, bone marrow, lymph node, and intestine. CCR7 is particularly expressed in lymphoid tissues and in activated B and T lymphocytes and has been shown to control the migration of memory T cells to inflamed tissues, as well as stimulate dendritic cell maturation. The chemokine (C-C motif) ligand 19 (CCL19/ECL) has been reported to be a specific ligand of this receptor. ESTs have been isolated from blood, embryo, lymph node, and thymus libraries. Receptors for the C - C chemokine family include CCR 1, CCR 2A, CCR 3, CCR 4, CCR 5 and the Duffy blood group antigen. The C-C receptors are important in the function of T cell chemotaxis and migration of phagocytic cells to sites of inflammation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: BLR2; Bukitt's lymphoma receptor 2; C-C chemokine receptor type 7; C-C CKR-7; CC chemokine receptor 7; CD197; CDw197; chemokine (C-C motif) receptor 7; EBI1; EBV-induced G protein-coupled receptor 1; Epstein-Barr virus induced gene 1; Epstein-Barr virus-induced G-protein coupled receptor 1; lymphocyte-specific G protein-coupled peptide receptor; MIP-3 beta receptor
Gene Aliases: BLR2; CC-CKR-7; CCR-7; CCR7; CD197; CDw197; CMKBR7; EBI1; EVI1
UniProt ID: (Human) P32248
Entrez Gene ID: (Human) 1236