Anti-AKT1 (phospho S473) antibody [EP2109Y]

• Supplier: Abcam
• Catalog: ab81283
• Prices: $404.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EP2109Y
• Applications: IHC-F IHC-P ICC/IF WB
• Species Reactivities: Mouse, Rat, Human
• Antigen: A phospho specific peptide corresponding to residues surrounding Ser473 of human AKT1
• Blocking Peptide: AKT1 peptide
• Description: Rabbit Monoclonal
• Gene: AKT1
• Conjugate: Unconjugated

Product images

NIH3T3 cells starved overnight and treated with PDGF 50ng/mL for 1 hour (A) or vehicle (B). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton-X100. Primary antibody ab81283 was used at 1:4000 dilution and secondary antibody Dylight GAR594 (ab96897) at 1:1000 dilution.

NIH3T3 cells were starved overnight and treated with PDGF 50ng/mL or vehicle control for 1 hour prior to fixation with 4% paraformaldehyde. Levels of total Akt were measured using antibody ab81283 on an infrared in cell ELISA assay platform.

Overlay histogram showing starved NIH3T3 cells treated with PDGF AB 50ng/mL for 1 hour (blue) and vehicle (red) stained with ab81283 antibody targeting Akt (phospho S473). Signal was analyzed by flow cytometry and compared with no primary antibody control (black). Cells were fixed with 4% paraformaldehyde for 20 minutes and permeabilized with methanol (30min) at -20C. Blocking was carred out in 5% BSA+0.05% tween in PBS. Primary antibody was diluted at 1:2000 and incubated for 1 hour at RT.

All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) at 1/10000 dilutionLane 1 : 3T3 cell lysate untreatedLane 2 : 3T3 cell lysates treated with PDGFLysates/proteins at 10 µg per lane.SecondaryHRP conjugated goat anti-rabbit at 1/2000 dilution

ab81283 staining AKT1 (phospho S473) in PC12 cells treated with galanin (1-29) (rat, mouse) (ab141153), by ICC/IF. Increase of AKT1 (phospho S473) expression correlates with increased concentration of galanin (1-29) (rat, mouse), as described in literature.The cells were incubated at 37°C for 24h in media containing different concentrations of ab141153 (galanin (1-29) (rat, mouse)) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81283 (1/100) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

ab81283, at 1/100 dilution, staining AKT1 in untreated (left panel) and Phosphatase-treated (right panel) cervical carcinoma by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue

Immunohistochemical analysis of Human HPV16 immortalized keratinocytes transfected with non-targeting siRNA, staining AKT1 (phospho S473) (green) with ab81283.Antigen retrieval was performed by heat mediation in citrate buffer (pH 6). Samples were blocked with 10% goat serum before incubating with primary antibody (1/100). Fluoroscein-conjugated tyramide was used to detect staining.

developed using the ECL techniquePerformed under reducing conditions.

ab81283 staining AKT1 (phospho S473) in PC3 cells treated with CAY10626 (ab120903), by ICC/IF. Decrease of AKT1 (phospho S473) expression correlates with increased concentration of CAY10626, as described in literature.The cells were incubated at 37°C for 24h in media containing different concentrations of ab120903 (CAY10626) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81283 (1/100) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

ab81283 staining AKT1 (phospho S473) in MCF7 cells treated with DAPT (ab120633), by ICC/IF. Decrease in expression of AKT1 (phospho S473) correlates with increased concentration of DAPT, as described in literature.The cells were incubated at 37°C for 24h in media containing different concentrations of ab120633 (DAPT) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81283 (1/200 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

developed using the ECL techniquePerformed under reducing conditions.

ab81283 staining AKT1 (phospho S473) in Human peritoneal tumor tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/500) for 2 hours. An Alexa Fluor® 647-conjugated Donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.See Abreview

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