AKT Multispecies In-Cell ELISA Kit, Colorimetric

Catalog number: 62215

Invitrogen™  Related applications: Protein Assays and Analysis

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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.

Features of the AKT Colorimetric In-Cell ELISA Kit:

• Measures total AKT and phospho-AKT (Ser473) in whole cells using 96-well or 384-well microplates
• 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

Two reliable detection formats are available:
• colorimetric detection kit for use with standard microplate readers
• near infrared fluorescent detection kit for use with IR imaging system

Overview of AKT and phosphorylation of AKT:
AKT proteins, also known 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 Products
Pierce™ AKT Near Infrared In-Cell ELISA Kit
For Research Use Only. Not for use in diagnostic procedures.


Assay Sensitivity: See product documentation
Detection Method: Colorimetric
Detector Antibody Conjugate: Unconjugated (uses Secondary Antibody)
For Use With (Equipment): Microplate Reader
Former Identifiers: Pierce
Gene ID: 10000, 207, 208
Gene Name: AKT serine/threonine kinase 1, AKT serine/threonine kinase 2, AKT serine/threonine kinase 3
Gene Symbol: AKT1, AKT2, AKT3
Immunoassay Kit Format: In-Cell ELISA Kit
Immunoassay Kit Grade: Colorimetric
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 96 tests
Protein Name: RAC-alpha serine/threonine-protein kinase, RAC-beta serine/threonine-protein kinase, RAC-gamma serine/threonine-protein kinase
Target (Kit Named): AKT
Target Species (Named): Multispecies
Target Species (Validated): Human
UniProt ID: P31749, P31751, Q9Y243

Contents & storage

Sufficient For: 96 microplate wells of overnight-cultured cells
• Blocking Buffer, 50 mL
• 20X Tris Buffered Saline, 50 mL
• Surfact-Amps 20 (10% Detergent), 10 mL
• Surfact-Amps X-100 (10% Detergent), 10 mL
• HRP Conjugate, 0.14 mL
• TMB Substrate, 58 mL
• TMB Stop Solution, 55 mL
• Janus Green Whole-Cell Stain, 50 mL
• Elution Buffer, 2 x 30 mL
• Thin Plate Seal Assembly, 8 each
• Anti-Phospho AKT (S473) Antibody
• Anti-AKT Antibody
Store antibodies as indicated on label. All other kit components should be stored at 4°C.


Manuals & protocols