Intracellular pH measurement with BCECF or pHrodo™ Red AM
Measurement of intracellular pH was performed using 5 µM of pHrodo™ Red AM Intracellular pH Indicator (Cat. No. P35372) or 5 µM of BCECF. pH clamping was carried out using a mixture of 10 mM nigericin and 10 mM valinomycin in Hepes-based buffers at different pHs. The fluorescence was measured using a high content screening (HCS) instrument. This data clearly indicates that while the signal of BCECF AM drops with drop in pH, signal of pHrodo™ Red AM increases with drop in pH. Further, while the signal of BCECF gets weaker due to photobleaching or other factors, the signal of pHrodo™ Red AM is consistent during this experiment.
Human peripheral blood leukocytes stained with CD45 FITC and antibody clone 581 PerCP-Cy®5.5 conjugate (top, Cat. No. A14949) or mouse IgG1,kappa PerCP-Cy®5.5 isotype control (bottom). Cy®togram gated for total live CD14 negative lymphocyte population. Go ›
Human peripheral blood leukocytes stained with CD34 antibody (clone 581) Pacific Blue conjugate (top, Cat. No. A14947) or mouse IgG1,kappa Pacific Blue isotype control (bottom) and costained with CD45 FITC, CD14 PE, and 7-AAD, then analyzed via gating on total live and CD14-negative lymphocyte population. Go ›