KeyTec® TR-FRET Phospho-AKT (S473) Kit

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Sandwich Model TR-FRET Assay for Detecting of P-AKT (S473) in Cell Lysates

• Easy to Use: A homogeneous assay - just follow the 'add-incubate-measure' process
• High Specificity: Use specific antibodies for binding P-AKT (S473), ensuring results accuracy
• Wider range for high concentration, making it more suitable for agonist or antagonistic (with agonist pre-treatment)

Sandwich Model TR-FRET Assay for Detecting of P-AKT (S473) in Cell Lysates

Size:

500 tests

5,000 tests

Price:

$ 2171

CAT.:

A1060031S

Delivery:

Available in Stock

Price:

$ 6312

CAT.:

A1060031L

Delivery:

Available in Stock

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KeyTec® TR-FRET Phospho-AKT (S473) Kit

500 tests

5,000 tests

CAT.: A1060031S

$ 2171 $ 6312

Overview

Performance

Protocols

Components

Notices

Overview

The KeyTec® TR-FRET Phospho-AKT (S473) Kit is designed for detect the level of phosphorylated AKT (S473) protein in cell lysates. It is based on sandwich immunoassay model and utilizes TR-FRET technology, known for its ease of use, homogeneity (no wash), low background, high sensitivity, robustness.
The detection principle is based on TR-FRET technology. A pair of anti Phospho-AKT (S473) protein antibodies, each labeled with KeyTec® TR-FRET Solar Eu and KeyTec® TR-FRET LA, bind to the same antigen Phospho-AKT (S473) protein. This binding brings the donor molecule into proximity with the acceptor molecule. Excitation of the donor will result in the generation of the TR-FRET signal at 665 nm, proportional to the concentration of Phospho-AKT (S473) protein.

Performance

Stimulation of Phospho-ERK1/2 (T202/Y204) in HEK293 cells
HEK293 cells (50K cells/well, in triplicate) were incubated with serial dilutions of EGF for 10 min at room temperature. The data show that treatment of HEK293 cells with EGF stimulates the phosphorylation of ERK1/2 at T202/Y204.

Inhibition of Phospho-ERK1/2 (T202/Y204) in HEK293 cells
HEK293 cells (50K cells/well, in triplicate) were incubated with serial dilutions of the inhibitor Erlotinib for 15 min at room temperature, followed by incubation with 0.5 nM EGF for 10 min at room temperature. The data show that treatment of HEK293 cells with Erlotinib inhibits the stimulated phosphorylation of ERK1/2 at T202/Y204.

Protocols

Protocol_A1060031_KeyTec® TR-FRET Phospho-AKT (S473) Kit_VKEYBIO-01-2024.pdf Download

Components

CAT.	Description	Size
A1060031S	KeyTec® TR-FRET Phospho-AKT (S473) Kit	500 tests
The tests are sufficient in a 384-well microplate assay format, with 20 μL per well. The details of the protocol refer to "KeyTec® TR-FRET Phosphorylated Protein Assay Kit General Guidebook". • 1 × 25 µL mAb anti-Phospho-AKT (S473) - Solar Eu • 1 × 100 µL mAb anti-Phospho-AKT (S473) - LA • 1 × 100 µL Positive control lysate • 1 × 250 µL Phosphatase Inhibitor Cocktail (100X) • 1 × 5 mL Phosphoproteins Lysis Buffer A (5X) • 1 × 250 µL Phosphoproteins Detection Buffer (10X)

Notices

Certificate of Analysis	Limitations	Storage Conditions
LOT.	For research use only	-60℃ or below

Overview

Performance

Protocols

Protocol_A1060031_KeyTec® TR-FRET Phospho-AKT (S473) Kit_VKEYBIO-01-2024.pdf Download

Components

CAT.	Description	Size
A1060031L	KeyTec® TR-FRET Phospho-AKT (S473) Kit	5,000 tests
The tests are sufficient in a 384-well microplate assay format, with 20 μL per well. The details of the protocol refer to "KeyTec® TR-FRET Phosphorylated Protein Assay Kit General Guidebook". • 1 × 25 µL mAb anti-Phospho-AKT (S473) - Solar Eu • 1 × 100 µL mAb anti-Phospho-AKT (S473) - LA • 1 × 100 µL Positive control lysate • 1 × 250 µL Phosphatase Inhibitor Cocktail (100X) • 1 × 5 mL Phosphoproteins Lysis Buffer A (5X) • 1 × 250 µL Phosphoproteins Detection Buffer (10X)

Notices

Certificate of Analysis	Limitations	Storage Conditions
LOT.	For research use only	-60℃ or below

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