PE Rat Anti-Human CCR7 (CD197)
Clone 3D12 (RUO)
- Brand BD Pharmingen™
- Alternative Name CCR7, BLR-2, EBI-1, CMKBR7
- Vol. Per Test 20 µl
- Isotype Rat IgG2a, κ
- Reactivity Human (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen Human CCR7 protein
- Entrez Gene ID 1236
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The monoclonal antibody 3D12 reacts with the human CC chemokine receptor, CCR7. CCR7 (previously known as BLR-2, EBI-1 and CMKBR7), a seven-transmembrane, G-protein-coupled receptor, is the specific receptor for CC chemokines, MIP-3β/Exodus 3/ELC/ CCL19 and 6Ckine/Exodus 2/SLC/TCA4/CCL21. It has been shown that CCR7 mRNA is expressed mainly in lymphoid tissues including spleen, lymph nodes and tonsil. CCR7 mRNA was also detected in peripheral T and B lymphocytes, in bone marrow and cord blood CD34-positive cells and mature dendritic cells. The human CCR7 gene, unlike other CC chemokine receptor genes, has been mapped to chromosome 17q12. The immunogen used to generate 3D12 hybridoma was the N-terminus as well as parts of the second extracellular loop of human CCR7 protein. The monoclonal antibody 3D12 recognizes an epitope mapping to the N-terminus of human CCR7.
- Format PE
- Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
- Excitation Max 496 nm
- Emission Max 578 nm
R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It exists in vitro as a 240-kDa protein with 23 phycoerythrobilin chromophores per molecule. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy.
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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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
The PE-conjugated 3D12 antibody is convenient for the immunofluorescent staining and flow cytometric analyses of human leukocytes and cell lines that express CCR7 (see figure). Chemokine receptors are known to internalize during manipulation resulting in low frequency expression. Immunophenotyping studies of chemokine receptors need to be performed on freshly collected whole blood (<24 hours). Incubation with the antibody should be done at 4°C in the dark. Cellular manipulation, such as Ficoll separation, freezing, or exposure to cold temperatures prior to staining have been shown to cause a decrease in staining intensity and inconsistent results.
A multiple-step staining procedure is strongly recommended to amplify immunofluorescent signals in the flow cytometric analysis of human CCR7 expression. Purified Rat Anti-Human CCR7 (CD197) (Cat. No. 552175) and a 3-step staining procedure is recommended for detecting low frequency expression.