AKT Activation InstantOne™ ELISA

Also known as: PKB, RAC-PK

RUO: For Research Use Only. Not for use in diagnostic procedures.

Attention: This 3 target InstantOne ELISAs contain reagents for a total of 96 wells (32 wells each for AKT (total), phospho-AKT (Thr308), and phospho-AKT (Ser473)). Please contact Technical Service for a lot specific COA

SKU# 85-86045

Cat. No. Size
85-86045-11 96 tests
Please inquire about custom configurations or bulk packaging.

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Data for AKT Activation InstantOne™ ELISA.

HEK293 cells were treated with either the PI3 Kinase inhibitor Wortmannin (2uM for 60 min) (-) or...View More

  • Data for AKT Activation InstantOne™ ELISA.
Description

Description: The InstantOne™ ELISA is specifically engineered for accurate measurement of human, mouse, and hamster total and phosphorylated AKT 1/2/3 (Thr308 and Ser473) in cell lysates. Detection of rat AKT 1/2/3 (Thr308 and Ser473) is also also expected. The InstantOne™ ELISA kit allows for fast analysis of samples in approximately one hour. All reagents used in a traditional sandwich ELISA are added in solution to a plate followed by a wash step and detection with the TMB colorimetric substrate.

AKT, which exists as multiple isoforms, is one of the principle kinases activated by phosphoinositide 3-kinase (PI3K). Activation of PI3K results in the generation of phosphatidylinositol (3,4,5)-triphosphate (PIP3). These lipid second messengers bind to the pleckstrin homology domain of AKT to promote its translocation to the plasma membrane for activation via phosphorylation at Thr308 and Ser473 by PDK1 and the mTOR TORC2 complex, respectively. Phosphorylation at both these sites is required for full activation of AKT Ser/Thr kinase activity. AKT phosphorylates over 50 known substrates, including GSK3, AS160, PRAS40, TSC 1, TCS 2, Raf-1, Bad, the FOXO family of transcription factors, and PFK2. Due to its established critical role in numerous biological processes, AKT is a frequent target for oncology-related drug development.

Details
Reactivity Human
Suitable Sample Types Tested on cell culture samples. Expected to work on many other sample types, but have not been tested at this time.
Components The kit includes sufficient reagents to perform 32 wells of each of the three specific targets for a total of 96 wells.
User Manual
InstantOne™ Assay Plate (Part No. IOAP96) - 96 well (sealed)
Cell Lysis Buffer (5X) (Part No. IOCLB1) - 8 mL
Enhancer Solution (Part No. IOES1) - 0.5 mL
Wash Buffer (10X) (Part No. IOWB1) - 8 mL
AKT 1/2/3 Capture Antibody Reagent (Part No. IOAKTACAR1T) - 1 mL
phospho-AKT 1/2/3 (Thr308) Capture Antibody Reagent (Part No. IOPAKTBCAR2T) - 1 mL
phospho-AKT 1/2/3 (Ser473) Capture Antibody Reagent (Part No. IOPAKTACAR1T) - 1 mL
AKT 1/2/3 Detection Antibody Reagent (Part No. IOAKTADAR1T) - 1 mL
phospho-AKT 1/2/3 (Thr308) Detection Antibody Reagent (Part No. IOPAKTBDAR2T) - 1 mL
phospho-AKT 1/2/3 (Ser473) Detection Antibody Reagent (Part No. IOPAKTADAR1T) - 1 mL
Positive Control Cell Lysate (lyophilized) (Part No. IOAKTAPC1) - 1 ea
Detection Reagent (TMB Substrate) (Part No. IODR1) - 10 mL
Stop Solution (Part No. IOSS1) - 10 mL
Plate Seal - 1 ea
Reported Applications ELISA
Documentation
TDS Link Download TDS
User Manual Download User Manual
Related Products
Cat. No. Name Excite Emit Application Reg.
85-86041 AKT 1/2/3 (Total) InstantOne™ ELISA ELISA RUO
85-86042 phospho-AKT 1/2/3 (Ser473) InstantOne™ ELISA ELISA RUO
85-86044 phospho-AKT 1/2/3 (Thr308) InstantOne™ ELISA ELISA RUO
85-86048 AKT Pathway Activation Profile InstantOne™ ELISA ELISA RUO
References

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