BD, 560922, BD Pharmingen™ PE-Cy™7 Rat Anti-Human CCR7 (CD197)

Alternative Name: CCR7, BLR-2, EBI-1, CMKBR7
Reactivity: Human (QC Testing)
Isotype: Rat IgG2a, κ
Immunogen: Human CCR7 protein
Application: Flow cytometry (Routinely Tested)
Vol. Per Test: 5 µl
RRID: AB_396765
Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: 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 PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.
Recommended Assay Procedures: Recommended Assay Procedures The Purified-, PE-, Biotin-, and PE-Cy7-conjugated 3D12 antibody (Cat No. 552175, 552176, 552174 and 557648) can be used for the immunofluorescent staining and flow cytometric analyses of human leukocytes and cell lines that express CCR7. Recommendation: 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 anti-human CCR7 (Cat. No. 552175) and a 3-step staining procedure is recommended for detecting low frequency expression.
Product Notices: Product Notices 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). For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036). Cy is a trademark of GE Healthcare. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.