BD, 559560, BD Pharmingen™ Purified Mouse Anti-Human CD196 (CCR6)

Alternative Name: BN-1; C-C CKR-6; C-C chemokine receptor type 6; CC-CKR-6; CCR-6
Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Isotype: Mouse IgG1, κ
Immunogen: Human CD196/CCR6 Peptide
Application: Flow cytometry (Routinely Tested)
Concentration: 0.5 mg/ml
Workshop Number: IX 48
RRID: AB_2291135
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 The purified 11A9 antibody (Cat. No. 559560) can be used for the immunofluorescent staining and flow cytometric analyses of human leukocytes  (see image) and cell lines that express CCR6. A multistep step staining procedure is recommended to amplify immunofluorescent signals for the flow cytometric analysis of human CCR6 expression: Step 1: Incubate the cells with 0.1 - 1 µg of purified 11A9 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 goat anti-mouse Ig (Cat. No. 553999) at 4°C for 20 minutes. Wash cells two times. Step 3: Incubate the cells with ≤ 0.06 mg of streptavidin-phycoerythrin (Cat. No. 554061) at 4°C for 20 minutes. Wash two times. Resuspend cells in staining medium and analyze stained cells with a BD FACScan™ Flow Cytometer (BD Biosciences, 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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.