Anti AKT, phospho Ser473  

Western Blot of Mouse anti-AKT pS473 antibody Atto 594 Conjugated. Lane 1: GST Tagged AKT 1 Active Recombinant Protein. Lane 2: None. Load: 25 ng per lane. Primary antibody: None. Secondary antibody: Atto 594 mouse secondary antibody at 1:1,000 for 60 min at RT. Block: MB-070 for 30 min at RT. Predicted/Observed size: ~100 kDa, ~100 kDa for AKT pS473. Other band(s): None

ATTO dyes can be used for multicolor immunofluorescent detection with low background and high signal. Examples shown are: A. Tubulin in PtK2- male Rat Kangaroo Kidney Epithelial Cells was detected using ATTO 532 labeled secondary antibody. B. Muscle alpha-actin was stained with a mouse primary antibody and ATTO 488 anti-mouse IgG (green) while Cytokeratin was stained with polyclonal rabbit anti-cytokeratin and ATTO 647N anti-rabbit IgG (red). C. HUVEC (Human umbilical vein endothelial cells were stained with anti- Vimentin-ATTO 532 (green), anti-E-Cadherin-ATTO 655 (red) and DAPI (blue). D. Rat colon sections were stained with Anti-Aquaporin 3-ATTO 594 antibody. Hoechst 33342 (blue) is used as counterstain.

Western Blot of Mouse Anti-Akt pS473 antibody. Lane 1: unstimulated NIH/3T3 lysates contain inactive unphosphorylated Akt1, green band. Lane 2: PDGF stimulated NIH/3T3 lysate contains both inactive (green band) and activated phosphorylated Akt1 (red band). Load: 10 μg per lane. Primary antibody: rabbit anti-Akt (pan) and mouse anti-Akt pS473 specific antibodies at 1:400 for overnight at 4°C. Secondary antibody: DyLight^TM 549 conjugated anti-rabbit IgG (green) and DyLight^TM 649 conjugated anti-mouse IgG (red) secondary antibodies at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C.