Akt (pS473) + total Akt ELISA Kit (ab126433)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Akt (pS473) + total Akt ELISA Kit -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay time
5h 00m -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Mouse, Rat, Human -
Product overview
ab126433 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphorylated Akt protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blot analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of phospho-Akt (Ser473) and total Akt in human, mouse and rat cell lysates (help normalize the results of phospho-Akt from different cell lysate being compared). A pan Akt antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Akt present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phospho-Akt (Ser473) or anti-pan-Akt is used to detect phosphorylated or total Akt. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Akt (Ser473) or total Akt bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Get higher sensitivity in only 90 minutes with AKT 1/2/3 pS473 + AKT1 ELISA Kit (ab176657) from our SimpleStep ELISA® range.
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Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate
Images
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Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)MCF7 cells were stimulated for five minutes with 1 ug x mL-1 of insulin (ab123768). Cell lysates were assessed for total Akt and Akt(pS473), shown as OD (450 nm) after background signal was subtracted (duplicates +/- SD).
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Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)The NIH3T3 cells were treated with recombinant human PDGF for 10 minutes to induce phosphorylation of Akt. Serial dilutions of lysates were analyzed in this ELISA. -
Western blot - Akt (pS473) + total Akt ELISA Kit (ab126433)NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed by Western Blot. -
Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed using this phosphoELISA. -
Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)A431 cells were treated with recombinant human EGF at 37°C for 20 min. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA.