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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.
|Components||1 x 96 tests|
|20X Wash Buffer Concentrate||1 x 25ml|
|2x Cell lysate buffer||1 x 5ml|
|5X Assay Diluent||1 x 15ml|
|Akt Microplate (12 strips x 8 wells) coated with anti-pan Akt antibody||1 unit|
|Detection Antibody Akt (Ser473): rabbit anti-phospho-Akt (Ser473)||1 vial|
|Detection Antibody Akt: rabbit anti-pan-Akt||1 vial|
|HRP-conjugated anti-rabbit IgG||1 x 25µl|
|Positive Control: lyophilized powder from NIH3T3 cell lysate||1 vial|
|Stop Solution||1 x 8ml|
|TMB Substrate Reagent||1 x 12ml|
Our Abpromise guarantee covers the use of ab126433 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"