Description
Staining Protocol: 1. Harvest cultured target cells into a 50 ml conical centrifuge tube. Centrifuge at 1000 rpm for 10 minutes, aspirate and discard supernate. 2. Wash cell pellet once with PBS and mix gently. Centrifuge at 1000 rpm for 10 minutes, aspirate and discard supernate. 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 supernate. 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 supernate. 5. Resuspend in PBS + 1% FBS (wash buffer) to a final concentration of approximately 1 x 10e6 per 50 µl. 6. Prepare one tube of 50 µl of cell suspension and add 20 µl of conjugated anti-human TdT. Prepare another tube of 50 µl of cell suspension and add 20 µl of conjugated isotype control. Shake gently, and incubate in the dark at room temperature for 20-30 minutes. 7. Wash tubes in 2 ml of wash buffer. Centrifuge for 5 minutes at 1000 rpm, aspirate and discard supernate. 8. Resuspend in 500 µl of wash buffer and analyze by flow cytometry.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
1. This antibody has been optimized and preassayed with its matched isotype control to be used at the recommended volume of 20 ul/test. Titration of the reagents or substituting with other (non-matched) isotype control is NOT recommended.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
4. 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.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
