BD, 552175, BD Pharmingen™ Purified 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)
Concentration: 0.5 mg/ml
RRID: AB_394353
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
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 3D12 antibody can be used for the immunofluorescent staining and flow cytometric analyses of human leukocytes and cell lines that express CCR7 (see Figure). A multiple-step staining procedure is strongly recommended to amplify immunofluorescent signals for the flow cytometric analysis of human CCR7 expression: 1. Incubate 10e6 cells with 0.1 - 0.5 µg of Purified Rat Anti-Human CCR7 (CD197) 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). 2. Incubate the cells with 0.25 µg of Biotin Mouse Anti-Rat IgG2a (Cat. No. 553894) at 4°C for 20 minutes. Wash cells two times. 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 instaining medium and analyze stained cells with a FACScan™ Flow Cytometer (BDIS, San Jose, CA) using appropriate specificity and compensation controls.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. An isotype control should be used at the same concentration as the antibody of interest. 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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use. 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.