BD, 560760, BD Phosflow™ T Cell Kit Lyophilized Cells

Application: Intracellular staining (flow cytometry) (Routinely Tested)
RRID: AB_2869365
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store unopened vials of lyophilized cells at 2 - 8°C. Do not freeze. Vials should be opened and the lyophilized cells reconstituted just prior to use.
Recommended Assay Procedures: Recommended Assay Procedures 1.  Gently tap the vial on a flat surface before using. This ensures that the lyophilized cell pellet is on the bottom of the vial. 2.  Open a vial of BD Phosflow™ Treated or Untreated Human Control Cells. Reconstitute the cells in 120 uL of BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) at room temperature. Gently vortex the cells for 1 to 2 seconds and let stand for at least 5 minutes before use. 3.  Transfer 100 µL of control cells to a 12 × 75-mm polystyrene tube. Keep cells on ice until ready to stain. Add fluorescent antibodies specific for intracellular signaling proteins and cell surface antigens (ie, fluorescent antibodies listed in the figure legend that have been validated for these cells), vortex the tube, and incubate at room temperature for 1 hour, protected from light. 4.  Wash cells by adding 2 mL of Stain Buffer and immediately centrifuging them  (600g for 6 min), aspirate the supernatant, and resuspend the stained cells in 300 to 500 µL of Stain Buffer. Analyze the cells by flow cytometry immediately (optimal) or no longer than 4 hours after preparation. Keep the cells at 2 to 8°C and protected from light prior to data acquisition.