BD, 550757, BD Pharmingen™ PE Mouse Anti-Human Terminal Transferase Set

Reactivity: Human (QC Testing)
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Regulatory Status: RUO
RRID: AB_393868
Description: Description Reacts with terminal deoxyribonucleotidyltransferase (TdT), a template-independent DNA polymerase that adds nucleotides to single stranded DNA primers. Western blot analysis of TdT reveals bands of 55, 40, and 15 kDa. TdT is found in normal bone marrow lymphoid progenitor cells and immature thymic lymphocytes. It has been reported that TdT is involved in the regulation and/or translocation of DNA and gene rearrangement during normal T and B cell development.
Preparation And Storage: Preparation And Storage 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.
Recommended Assay Procedures: Recommended Assay Procedures 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.