Akt (pS473) + total Akt ELISA Kit (ab126433)
- Product nameAkt (pS473) + total Akt ELISA KitSee all AKT 1/2/3 (phospho Ser473) + total AKT 1/2/3 kits ...
- Detection methodColorimetric
- Tests1 x 96 well plate
- Sample typeCell Lysate
- Assay typeSandwich
- Assay time5h 00m
- Assay durationMultiple steps standard assay
- Species reactivityReacts 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.
- Tested applicationsSandwich ELISA more details
- Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 x 96 tests 20X Wash Buffer Concentrate 1 x 25ml 2X Cell Lysate Buffer 1 x 5ml Akt Microplate (12 strips x 8 wells) coated with anti-pan Akt antibody 1 unit Assay Diluent: 5x concentrated buffer 1 x 15ml 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: 1000x HRP-conjugated anti-rabbit IgG concentrate 1 x 25µl Positive Control: lyophilized powder from A431 cell lysate 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml
- Research Areas
Contains 1 AGC-kinase C-terminal domain.
Contains 1 PH domain.
Contains 1 protein kinase domain.
modificationsPhosphorylation on Thr-309 and Ser-474 is required for full activity.
Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT2 ubiquitination. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome.
- AKT2_HUMANAKT3HIHGHHPKBPKB alphaPKB betaPKB gammaPKB-ALPHAPKB-GAMMAPKBBPKBBETAPKBGPRKBAPRKBBPRKBGProtein kinase Akt-2Protein kinase Akt-3Protein kinase BProtein kinase B alphaProtein kinase B betaProtein kinase B gammaProto oncogene c AktRACRAC-ALPHARAC-alpha serine/threonine-protein kinaseRAC-BETARAC-beta serine/threonine-protein kinaseRAC-gammaRAC-gamma serine/threonine-protein kinaseRAC-PK-alphaRAC-PK-betaRAC-PK-gammaSTK-2V-akt murine thymoma viral oncogene homolog 1V-akt murine thymoma viral oncogene homolog 2V-akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma)
- Entrez Gene: 208 Human
- Entrez Gene: 10000 Human
- Entrez Gene: 207 Human
- Entrez Gene: 11652 Mouse
- Entrez Gene: 11651 Mouse
- Entrez Gene: 23797 Mouse
- Entrez Gene: 25233 Rat
- Entrez Gene: 24185 Rat
- Entrez Gene: 29414 Rat
- Omim: 164731 Human
- Omim: 164730 Human
- Omim: 611223 Human
- SwissProt: P31751 Human
- SwissProt: P31749 Human
- SwissProt: Q9Y243 Human
- SwissProt: Q60823 Mouse
- SwissProt: P31750 Mouse
- SwissProt: Q9WUA6 Mouse
- SwissProt: P47197 Rat
- SwissProt: P47196 Rat
- SwissProt: Q63484 Rat
- Unigene: 631535 Human
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.
Akt (pS473) + total Akt ELISA Kit images
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.
NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed using this phosphoELISA.
NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed by Western Blot.
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.
References for Akt (pS473) + total Akt ELISA Kit (ab126433)
ab126433 has not yet been referenced specifically in any publications.