Western blot analysis of Nur77 expression on mouse thymocytes. Thymocytes were stimulated with PMA (20 ng/ml) and ionomycin (500 ng/ml at 37°C for 2 hr). Lysates were prepared and separated by SDS/PAGE. Blots were probed with Purified Mouse Anti-Nur77 (Cat. No. 554088) at concentrations of 2.0 (lane 1), 1.0 (lane 2), and 0.5 µg/ml (lane 3). Visualization was carried out with HRP Goat Anti-Mouse Ig (Cat. No. 554002). Nur77 is detected as a protein of ~77 kDa.

Left Figure: Western blot analysis of Nur77 expression on mouse thymocytes. Thymocytes were stimulated with PMA (20 ng/ml) and ionomycin (500 ng/ml at 37°C for 2 hr). Lysates were prepared and separated by SDS/PAGE. Blots were probed with Purified Mouse Anti-Nur77 (Cat. No. 554088) at concentrations of 2.0 (lane 1), 1.0 (lane 2), and 0.5 µg/ml (lane 3). Visualization was carried out with HRP Goat Anti-Mouse Ig (Cat. No. 554002). Nur77 is detected as a protein of ~77 kDa. Right Figure: Flow cytometric analysis of Nur77 expression on stimulated mouse thymocytes. Thymocytes from C57/BL/6 mice were left unstimulated (gray line histogram) or stimulated (black line histograms) with PMA (20ng/ml) and  Ionomycin (500ng/ml) for 2 hours and then stained intracellularly with Purified Mouse IgG1, k Isotype Control (Cat.No. 554121; black dashed line) or Purified Mouse Anti-Nur77 antibody (black solid line) at 0.125ug/test using BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574). Purified antibody was followed by PE conjugated Goat Anti-Mouse Ig (Multiple Absorption Secondary antibody (Cat.No. 550589). Histogram plots were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.