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 Staining Buffer (Product No. SB-4400) 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 No. 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 No. 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 No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizationsregarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser Apoptosis is induced by certain cytokines including TNF and Fas ligand in the TNF family through their death domain containing receptors. TRAIL/Apo2L is a new member of the TNF family and induces apoptosis of a variety of tumor cell lines. DR4 and DR5 are the recently identified functional receptors for TRAIL, and DcR1/TRID is a decoy receptor. Another member of the TRAIL receptor family was more recently identified and designated DcR2, TRAIL-R4, or TRUNDD. DcR2 has an extracellular TRAIL-binding domain but lacks intracellular death domain and does not induce apoptosis. Like DR4 and DR5, DcR2 transcript is widely expressed in normal human tissues. Overexpression of DcR2 attenuated TRAIL-induced apoptosis.
|4° C, store in dark, DO NOT FREEZE!|
|Super Bright 780|
|PBS with BSA and 0.09% sodium azide; pH 7.2|