BD, 550937, BD Pharmingen™ Purified Mouse Anti-Human CD197 (CCR7)

Alternative Name: CCR7, BLR-2, EBI-1 and CMKBR7
Reactivity: Human (QC Testing)
Isotype: Mouse IgM, κ
Immunogen: Human CCR7 transfected L1.2 mouse lymphoma cells
Application: Flow cytometry (Routinely Tested)
Concentration: 0.5 mg/ml
RRID: AB_393968
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
Recommended Assay Procedures: Recommended Assay Procedures 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. The purified 2H4 antibody can be used for the immunofluorescent staining and flow cytometric analyses of human leukocytes and cell lines that express CCR7 (see Figure). A multistep step staining procedure is recommended to amplify immunofluorescent signals for the flow cytometric analysis of human CCR7 expression: Step 1: Incubate 10e6 cells with 2 - 4 µg of purified 2H4 antibody at 4°C for 15 - 20 minutes. Wash cells two times with staining medium containing sodium azide (e.g., Dulbecco's PBS or tissue culture medium [without phenol red and biotin] with 0.09% sodium azide and 2% heat-inactivated FCS or 0.2% BSA). Step 2: Incubate the cells with 0.25 µg of biotinylated rat anti-mouse IgM (Cat. No. 553406) at 4°C for 20 minutes. Wash cells two times. Step 3: Incubate the cells with ≤ 0.06 µg of streptavidin-phycoerythrin (Cat. No. 554061) at 4°C for 20 minutes. Wash two times. Resuspend cells in staining medium and analyze by flow cytometry using appropriate specificity and compensation controls.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols. 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.