BD, 552174, BD Pharmingen™ Biotin 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: 20 µl
RRID: AB_394352
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 biotin under optimum conditions, and unreacted biotin was removed.
Recommended Assay Procedures: Recommended Assay Procedures The biotinylated 3D12 antibody can be used with a very bright second-step reagent, such as Streptavidin-phycoerythrin (Cat. No. 554061) for the immunofluorescent staining and flow cytometric analyses of human leukocytes and cell lines that express CCR7 (see Figure). We are unable to generate reproducible results using less bright reagent such as Streptavidin-FITC or -PerCP with this antibody. The use of Streptavidin-FITC or -PerCP is not recommended. Recommendation: Chemokine receptors are know to internalize during manipulation resulting in low frequency expression. Immunophenotyping studies of chemokine receptors need to be performed on freshly collected whole blood (<24 Hrs). Incubation with the antibody should be done 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 CD197 (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). 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.