Anti-Notch1 Antibody [S253-32] (A304795)

Immunocytochemistry/Immunofluorescence analysis of human neuroblastoma cells (SH-SY5Y), fixed in 4% PFA for 15 min, using Anti-Notch1 Antibody [S253-32] (A304795), at 1:50 for overnight at 4°C with slow rocking. The secondary antibody used was AlexaFluor 488 at 1:1,000 for 1 hour at room temperature. Counterstain: Phalloidin-iFluor 647 (red) F-Actin stain; Hoechst (blue) nuclear stain at 1:800, 1.6mM for 20 minutes at room temperature.(A) Hoechst (blue) nuclear stain. (B) Phalloidin-iFluor 647 (red) F-Actin stain. (C) Notch1 Antibody (D) Composite.

Western blot analysis of rat brain membrane showing detection of ~270 kDa Notch1 protein using Anti-Notch1 Antibody [S253-32] (A304795) at 1:1,000 for 1 hour at room temperature. Lane 1: MW Ladder. Lane 2: rat brain membrane (10 µg).. Load: 10 µg. Block: 5% milk. The secondary antibody used was Goat Anti-Mouse IgG: HRP at 1:200 for 1 hour at room temperature. Color Development: TMB solution for 10 min at room temperature. Predicted/Observed Size: ~270 kDa.

Immunocytochemistry/Immunofluorescence analysis of human neuroblastoma cell line (SK-N-BE, fixed in 4% formaldehyde for 15 min at room temperature, using Anti-Notch1 Antibody [S253-32] (A304795), at 1:100 for 60 minutes at room temperature. The secondary antibody used was Goat Anti-Mouse ATTO 488 at 1:100 for 60 minutes at room temperature. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5,000 for 60min room temperature, 5min room temperature. Localization: Cell Membrane, Nucleus. Magnification: 60X.(A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) Notch1 Antibody. (D) Composite.