Rabbit Anti-AKTpS473 Antibody was prepared by repeated immunizations in rabbits with a synthetic peptide corresponding to a C-terminus region near phospho Serine 473 of the human, mouse, rat and chicken AKT proteins conjugated to KLH.
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Formulierung:
Liquid (sterile filtered)
Target-Kategorie:
Human
Antibody Type:
Primary Antibody
Application Verdünnung:
ELISA: 1:15,000 - 1:60,000, IHC: 1:100 - 1:500, IF Microscopy: User Optimized, WB: 1:200 - 1:1000
Anwendungsbeschreibung:
This AKT antibody is phospho specific for pS473 and is tested in ELISA, western blotting, immunohistochemistry (formalin-fixed paraffin-embedded sections), and immunofluorescence. By immunoblot a single band of the expected apparent molecular weight ~56k
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: Dakos 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 2-8C. 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 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: Dakos 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: Dakos 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 4C. 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.
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