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.