Anti-AKT1 (phospho S473) antibody (ab18206)
Key features and details
- Rabbit polyclonal to AKT1 (phospho S473)
- Suitable for: IHC-P, WB
- Reacts with: Mouse
- Isotype: IgG
Overview
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Product name
Anti-AKT1 (phospho S473) antibody
See all AKT1 primary antibodies -
Description
Rabbit polyclonal to AKT1 (phospho S473) -
Host species
Rabbit -
Specificity
The immunogen was raised against Human AKT1 phosphorylated at serine 473, however due to the high degree of similarity between AKT1, AKT2 and AKT3 the antibody is also predicted to react with all these isoforms. -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse
Predicted to work with: Rat, Cow, Human, Xenopus laevis, African green monkey
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab18622) -
Positive control
- This antibody gave a positive signal in NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate IHC-P: FFPE mouse lung tissue sections.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Images
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Western blot - Anti-AKT1 (phospho S473) antibody (ab18206)All lanes : Anti-AKT1 (phospho S473) antibody (ab18206) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate
Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate withHuman pan-AKT (phospho S473) peptide (ab18622) at 1 µg/ml
Lane 4 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate withHuman pan-AKT (phospho S473) peptide (ab18622) at 1 µg/ml
Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Human AKT1 (unmodified ) peptide (ab24649) at 1 µg/ml
Lane 6 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate with Human AKT1 (unmodified ) peptide (ab24649) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 57-60 kDa why is the actual band size different from the predicted?
Additional bands at: 130 kDa, 78 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
We are unsure of the identity of the higher bands detected at ~78 and 130 kDa. We believe this could be cross reaction with other phospho proteins, however the target band at 57 kDa is detected only in PDGF treated lysate and this signal is quenched by addition of the immunising peptide suggesting a specific interaction. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody (ab18206)IHC image of AKT1 (phospho S473) staining in mouse lung formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18206, 5µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
