BD, 558257, BD Pharmingen™ APC Mouse Anti-Human CD3ε

Reactivity: Human (QC Testing), Mouse (Reported)
Isotype: Mouse IgG1, κ
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Vol. Per Test: 20 µl
RRID: AB_647163
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 to APC under optimum conditions, and unconjugated antibody and free APC were removed.
Recommended Assay Procedures: Recommended Assay Procedures Protocol for PBMC cells fixation, permeabilization and staining 1. Harvest, count and pellet PBMC cells following standard procedures. 2. While vortexing, add 5 ml cold 70% - 80% ethanol dropwise into the cell pellet (1-5 x 10^7 cells). Incubate at -20°C for at least 2 hours. These fixed cells can be stored at -20°C for up to 60 days prior to staining. 3. Wash twice with 30-40 ml 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/ml. 5. Transfer 100 µl (1 x 10^6 cells) cell suspension into each sample tube. 6. Add 20 µl of properly diluted fluorescence conjugated antibody into the tubes above. Mix gently. 7. Incubate the tubes at room temperature (RT) for 20-30 minutes in the dark. 8. Wash with 2 ml of staining buffer at 200 x g for 5 minutes. 9. Aspirate the supernatant. 10. Add 0.5 ml of staining buffer to each tube. 11. Proceed to flow cytometric analysis.
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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser. Please refer to http://regdocs.bd.com to access safety data sheets (SDS). Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.