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AKT (Total/Phospho) InstantOne™ ELISA

Also known as: Protein Kinase B, PKB, RAC-alpha serine/threonine-protein kinase

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

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SKU# 85-86046

Cat. No. Size
85-86046-11 96 tests

Data for AKT (Total/Phospho) InstantOne™ ELISA.

MCF7 cells were pre-treated with 10 uM wortmannin washed and unstimulated (-) or stimulated with 10...View More

  • Data for AKT (Total/Phospho) InstantOne™ ELISA.
  • Data for AKT (Total/Phospho) InstantOne™ ELISA.
  • Data for AKT (Total/Phospho) InstantOne™ ELISA.

Description: The InstantOne™ ELISA is specifically engineered for accurate measurement of human, mouse, and hamster total AKT 1 and phosphorylated AKT 1/2/3 (Ser473) in cell lysates. Detection of rat AKT 1 is 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.

Reactivity Human, Mouse, Rat
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 AKT (Total/Phospho) InstantOne™ ELISA contains sufficient reagents to detect a total of 96 wells (48 wells of total AKT and 48 wells of phospho-AKT)
1. User Manual
2. InstantOne™ Assay Plate (Part No. IOAP96) - 96 well (sealed)
3. Cell Lysis Buffer (5X) (Part No. IOCLB1) - 8 mL
4. Enhancer Solution (Part No. IOES1) - 0.5 mL
5. Wash Buffer (10X) (Part No. IOWB1) - 8 mL
6. AKT 1 (Total) Capture Antibody Reagent (Part No. IOAKTBCAR1M) – 1 x 0.75 mL
7. phospho-AKT 1/2/3 (Ser473) Capture Antibody Reagent (Part No. IOPAKTACAR1M) – 3 x 0.75 mL
8. AKT 1 (Total) Detection Antibody Reagent (Part No. IOAKTBDAR1M) – 1 x 0.75 mL
9. phospho-AKT (Ser473) Detection Antibody Reagent (Part No. IOPAKTADAR1M) – 3 x 0.75 mL
10. Positive Control Cell Lysate (lyophilized) (Part No. IOAKTAPC1) - 1 ea
11. Detection Reagent (TMB Substrate) (Part No. IODR1) - 10 mL
12. Stop Solution (Part No. IOSS1) - 10 mL
13. Plate Seal - 1 ea
Volume Reagents for 24 wells of total AKT and 72 wells of phospho-AKT (Ser473)
Reported Applications ELISA
TDS Link Download TDS
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85-86018 ERK/AKT/p70 S6K Activation InstantOne™ ELISA ELISA RUO
85-86047 AKT 1 (Total) InstantOne™ ELISA ELISA RUO
85-86014 AKT/ERK Activation InstantOne™ ELISA ELISA RUO
85-86042 phospho-AKT 1/2/3 (Ser473) InstantOne™ ELISA ELISA RUO

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