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AKT In-Cell ELISA Kits


Microplate assays to measure total and phospho-AKT in whole cells.

Pierce AKT In-Cell ELISA Kits

Thermo Scientific Pierce AKT In-Cell ELISA Kits are colorimetric and fluorescent microplate assays optimized to measure total and phosphorylated Akt family of serine/threonine-specific protein kinases (phospho-AKT) in whole cells.

These ELISA kits for whole cells include a mouse monoclonal anti-AKT antibody, rabbit monoclonal anti-phospho-(Ser473) AKT antibody, detection reagents and all buffers required for either colorimetric or near infrared detection. The AKT In-Cell ELISA Kits enable simultaneous detection of both total and phospho-(Ser473) AKT.

Highlights:

  • Measures total AKT and phospho-AKT (Ser473) in whole cells using 96-well or 384-well microplates
  • Two reliable detection formats available:
    • colorimetric detection kit for use with standard microplate readers
    • near infrared fluorescent detection kit for use with IR imaging system
  • Whole-cell detection requires no cell lysis, protein extraction or special sample preparation
  • Provides better data and higher throughput than quantitative Western blotting
  • Less costly (one well vs. one lane) than Western blotting

Product Details:

Stimulation with EGF causes increased AKT phosphorylation A431 cells. Relative total and phospho-AKT levels were measured in A431 cells using the AKT (A) Colorimetric and (B) Near IR In-Cell ELISA Kits. Cells were serum-starved overnight and stimulated with 100ng/ml EFG for five minutes immediately prior to fixation. Total AKT was not significantly different between experiments or treatments. Note: Absolute absorbance values cannot be directly compared between phospho and total AKT experiments because they use antibodies with different sensitivities.

Overview of AKT and phosphorylation of AKT:

AKT proteins, also know as protein kinases B (PKB), are important for cell signaling in mammals. AKT1, 2, and 3 are serine/threonine-specific kinases. AKT1 is involved in cell survival and inhibition of apoptosis and is implicated in human cancers as well as growth regulation in mice. AKT2 has role in the insulin signaling pathway and has been shown to be linked to a diabetic phenotype in mice. AKT3 is highly expressed in the brain and knock-out of AKT3 results in reduced brain mass in mice. AKT proteins have a PH domain which binds to phosphoinositides which are phosphorylated by PI3-kinases. When located at the cell membrane and bound to PIP3, AKT protein can be phosphorylated by mTORC2 (Ser 473) allowing subsequent phosphorylation by PDPK1 (Thr 308). This activates AKT kinase activity which leads to the regulation of downstream targets. The AKT In-Cell ELISA Kits enable simultaneous detection of both total and phospho-(Ser473) AKT.

Related Resources:

Overview and Menu of Target-Specific In-Cell ELISA Kits
Dylight Fluor Spectral Properties (Ex/Em)

Related Products:

ELISA reagent reservoirs and sealing tape
Secondary Antibodies and Conjugates For ELISAs and Western Blotting
DyLight Fluor Conjugated Secondary Antibodies and Biotin Binding Proteins

Notice:

This product is for In Vitro experimental use only. Not for resale without express authorization.


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
62215 AKT Colorimetric In-Cell ELISA Kit
Formulation: Multi-component kit
Sufficient For: 96 microplate wells of overnight-cultured cells

Kit Contents:
Blocking Buffer, 50mL
20X Tris Buffered Saline, 50mL
Surfact-Amps 20 (10% Detergent), 10mL
Surfact-Amps X-100 (10% Detergent), 10mL
HRP Conjugate, 0.14mL
TMB Substrate, 58mL
TMB Stop Solution, 55mL
Janus Green Whole-Cell Stain, 50mL
Elution Buffer, 2 x 30mL
Thin Plate Seal Assembly, 8 each
Anti-Phospho AKT (S473) Antibody
Anti-AKT Antibody
96-test kit Product Instructions for product #62215 AKT Colorimetric In-Cell ELISA Kit MSDS for product #62215 AKT Colorimetric In-Cell ELISA Kit $380.00

Add to cart
62220 AKT Near Infrared In-Cell ELISA Kit
Formulation: Multi-component kit
Sufficient For: 96 microplate wells of overnight-cultured cells

Kit Contents:
Blocking Buffer, 100mL
20X Tris Buffered Saline, 50mL
Surfact-Amps 20 (10% Detergent), 10mL
Surfact-Amps X-100 (10% Detergent), 10mL
DyLight 680 Goat Anti-Mouse IgG (H+L), 55µL
DyLight 800 Goat Anti-Rabbit IgG (H+L), 55µL
Thin Plate Seal Assembly, 8 each
Anti-Phospho AKT (S473) Antibody
Anti-AKT Antibody
96-test kit Product Instructions for product #62220 AKT Near Infrared In-Cell ELISA Kit MSDS for product #62220 AKT Near Infrared In-Cell ELISA Kit $380.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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