Phospho-AKT / GSK3 beta / mTor ELISA kit (ab279732)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Mouse, Human
Overview
-
Product name
Phospho-AKT / GSK3 beta / mTor ELISA kit -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Mouse, Human -
Product overview
Phospho-AKT / GSK3 beta / mTor ELISA Kit (ab279732) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human and mouse cell lysates. By determining phosphorylated AKT/ GSK3 beta / mTor 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 Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human and mouse phospho-AKT/ GSK3 beta / mTor and total-AKT/ GSK3 beta / mTor. For each target, a capture antibody has been coated onto microwells. Samples are pipetted into the wells and target protein present in a sample is bound to the wells by the immobilized antibody. The wells are washed, and a detection antibody is used to detect the captured target protein. After washing away unbound antibody, an HRP-conjugated secondary antibody 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 target protein bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
-
Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
-
Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 100X HRP-Streptavidin Concentrate 1 x 200µl 20X Wash Buffer Concentrate 1 x 25ml 2X Cell lysate buffer 1 x 10ml 5X Assay Diluent 1 x 15ml Biotinylated Rabbit anti-mTOR antibody 1 vial HRP-conjugated anti-rabbit IgG concentrate (1000X) 1 vial Pan-AKT / GSK3 beta / mTor Coated Microplate 1 unit Positive Control - NIH/3T3 cell lysate 1 vial Rabbit anti-AKT antibody 1 vial Rabbit anti-GSK beta antibody 1 vial Rabbit anti-phospho-AKT (S473) antibody 1 vial Rabbit anti-phospho-GSK beta (S9) antibody 1 vial Rabbit anti-phospho-mTOR (S2448) antibody 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
Cellular localization
mTOR: Endoplasmic reticulum membrane. Golgi apparatus membrane. Mitochondrion outer membrane. Lysosome. Cytoplasm. Nucleus > PML body. Shuttles between cytoplasm and nucleus. Accumulates in the nucleus in response to hypoxia (By similarity). Targeting to lysosomes depends on amino acid availability and RRAGA and RRAGB. GSK3 beta: Cytoplasm. Nucleus. Cell membrane. The phosphorylated form shows localization to cytoplasm and cell membrane. The MEMO1-RHOA-DIAPH1 signaling pathway controls localization of the phosophorylated form to the cell membrane. AKT1: Cytoplasm. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus. -
Database links
- Entrez Gene: 207 Human
- Entrez Gene: 2475 Human
- Entrez Gene: 2932 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 56637 Mouse
- Entrez Gene: 56717 Mouse
- Omim: 164730 Human
- Omim: 601231 Human
see all
Images
-
NIH/3T3 cells were treated with recombinant human PDGFBB at 37oC for 10 mins. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA.
-
A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 20 mins.
Cell lysates were analyzed using ELISA and Western Blot.
-
NIH/3T3 cells were treated or untreated with 50 ng/ml recombinant human PDGFBB for 10 min.
-
NIH/3T3 cells were treated or untreated with 50 ng/ml recombinant human PDGFBB for 10 min.
-