Anti AKT, phospho Ser473  

Immunohistochemistry of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.

Dot Blot of Rabbit Anti-AKT pS473 Antibody. Dilutions in Columns: (1) 100ng, (2) 33.33ng, (3) 11.11ng, (4) 3.7ng, (5) 1.23ng. Tested BSA Peptide Reactivity in Rows: (A) AKT1-BSA, (B) AKT1 pT308-BSA, (C) AKT1 S473-BSA, (D) AKT1 pS473-BSA, (E) CDC27 T244-BSA, (F) CDC27 pT244-BSA, (G) BSA control. Primary Antibody: Anti-AKTpS473 at 1μg/mL overnight at 2-8°C. Secondary Antibody: Goat anti-Rabbit IgG HRP (p/n 611-103-122) at 1:70,000 at RT for 30mins. Block: BlockOut Buffer (p/n MB-073).

Immunohistochemistry at higher magnification of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.

Immunofluorescence Confocal Microscopy of Rabbit anti-AKT pS473 antibody. Tissue: cardiomyocytes infected with adenovirus expressing with wild-type AKT. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 1:40 for 1 h at RT. Secondary antibody: texas-red conjugated rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: AKT pS473 is nuclear. Staining: AKT pS473 as green fluorescent signal with texas-red conjugated phalloidin (red) to label filamentous actin.

Immunohistochemistry of Rabbit anti-AKT pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 1:100 for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: AKT pS473 is nuclear and occasionally cytoplasmic.

Western Blot of Rabbit anti-AKT pS473 antibody. Lane 1: AKT1 Recombinant Protein (p/n 009-001-P21). Lane 2: AKT1 Mutant Human Recombinant Protein (p/n 009-001-P22). Lane 3: AKT1 (phosphatase treated) Human Recombinant Protein (p/n 009-001-I51). Load: 50 ng per lane. Primary antibody: AKT pS473 antibody at 1:1,000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n 611-103-122) at 1:40,000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting (MB-070) for 30 min at RT. Predicted/Observed size: ~56 kDa for AKTpS473.