BD, 563755, BD Horizon™ BV711 Mouse Anti-Ki-67

Alternative Name: MKI67; Antigen identified by monoclonal antibody Ki-67; KIA
Reactivity: Human (QC Testing), Mouse (Tested in Development), Rat, Rhesus (Reported)
Isotype: Mouse IgG1, κ
Immunogen: Human Ki-67
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Vol. Per Test: 5 µl
RRID: AB_2738406
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 BD Horizon™ BV711 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV711 were removed.
Recommended Assay Procedures: Recommended Assay Procedures Investigators are encouraged to utilize the following protocol with the use of DAPI.  Higher background or signal spillover may be experienced with the use of 7-AAD or propidium iodide (PI). 1. Harvest, count and pellet cells following standard procedures. Note: Ki-67 is expressed by proliferating cells. Using resting cells (e.g, unstimulated PBMC) may give negative results. 2. While vortexing, add 5 mL cold 70% ethanol dropwise into the cell pellet (1-5 x 10^7 cells). 3. Wash twice with staining buffer (PBS with 1% FBS, 0.09% NaN3), centrifuge for 10 minutes at 200 x g. 4. Resuspend the cells to a concentration of 1 x 10^7 cells/mL. 5. Transfer 100 µL (1 x 10^6 cells) cell suspension into each sample tube. 6. Add 5 µL of  BV711 Mouse Anti-Ki-67 antibody into the appropriates tubes. Mix gently. 7. Incubate the tubes at 4°C for 60 min in the dark. 8. Wash with 2 mL of staining buffer at 200 x g for 5 minutes. 9. Aspirate the supernatant. 10. Repeat wash with 2 mL of staining buffer at 200 x g for 5 minutes. 11. Aspirate the supernatant. 12. Resuspend in 350-500 µL DAPI (Cat# 564907) diluted to 1 µg/mL. 13. Proceed to flow cytometric analysis. For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application. Cy is a trademark of GE Healthcare. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR. BD Horizon Brilliant Violet 711 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239. 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.