BD, 565230, BD Horizon™ BV421 Mouse Anti-Human Terminal Transferase (TdT)

Alternative Name: TdT; DNTT; Terminal transferase; Terminal addition enzyme
Reactivity: Human (QC Testing)
Isotype: Mouse BALB/c IgG1, κ
Immunogen: Purified Human TdT
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Vol. Per Test: 5 µl
RRID: AB_2739124
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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
Recommended Assay Procedures: Recommended Assay Procedures 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). Staining Protocol: 1. Harvest cultured target cells into a 50 ml conical centrifuge tube. Centrifuge at 1000 rpm for 10 minutes, aspirate and discard supernatant. 2. Wash cell pellet once with PBS and mix gently. Centrifuge at 1000 rpm for 10 minutes, aspirate and discard supernatant. 3. Fix the cells by adding 15-20 ml of 1% formaldehyde while vortexing the pellet and incubate for 20 minutes at room temperature. Centrifuge at 1000 rpm for 10 minutes, aspirate and discard supernatant. 4. Add 15-20 ml of 0.1% Triton™ X-100 in PBS and incubate for 5-10 minutes. Centrifuge at 1000 rpm for 10 minutes, aspirate and discard supernatant. 5. Resuspend cells in PBS + 1% FBS (wash buffer) to a final concentration of approximately 1 x 10^6 cells per 50 µl. 6. Prepare one tube with 50 µl of cell suspension and add 20 µl of fluorescent Anti-Human TdT antibody. Prepare another tube with 50 µl of cell suspension and add 20 µl of a matched fluorescent Ig isotype control. Mix the tube contents gently, and incubate in the dark at room temperature for 20-30 minutes. 7. Wash cells with 2 ml of wash buffer per tube. Centrifuge for 5 minutes at 1000 rpm, aspirate and discard supernatant. 8. Resuspend cells in 500 µl of wash buffer and analyze by flow cytometry.
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). Please refer to www.bdbiosciences.com/us/s/resources for technical protocols. Triton is a trademark of the Dow Chemical Company. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR. 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. An isotype control should be used at the same concentration as the antibody of interest.