AKT (Phospho) [pT308] Multispecies InstantOne™ ELISA Kit

Catalog number: 85-86044-11

Invitrogen™  Related applications: Protein Assays and Analysis

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Description

The InstantOne™ ELISA is specifically engineered for accurate measurement of phosphorylated human, mouse, and hamster AKT 1/2/3 (Thr308) in cell lysates. Detection of rat AKT 1/2/3 (Thr308) is 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.

Suitable Sample Types
Tested on cell culture samples. Expected to work on many other sample types, but have not been tested at this time.

Reported Application
ELISA

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

Specifications

Assay Range: See product documentation
Assay Sensitivity: See product documentation
Detection Method: Colorimetric
For Use With (Equipment): Microplate Reader
Former Identifiers: eBioscience
Gene Aliases: AKT, CWS6, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA, HIHGHH, PKBB, PKBBETA, PRKBB, RAC-BETA, MPPH, MPPH2, PKB-GAMMA, PKBG, PRKBG, RAC-PK-gamma, RAC-gamma, STK-2
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: Coated ELISA Kit
Label or Dye: HRP (Horseradish Peroxidase)
Product Line: InstantOne™
Product Size: 96 tests
Protein Name: RAC-alpha serine/threonine-protein kinase, RAC-beta serine/threonine-protein kinase, RAC-gamma serine/threonine-protein kinase
Sample Type (Specific): Cell Extracts
Target (Kit Named): AKT
Target Site-Specificity: [pT308]
Target Species (Named): Multispecies
Target Species (Validated): Human
Target Sub-Specificity: Phospho
UniProt ID: P31749, P31751, Q9Y243

Contents & storage

The kit includes sufficient reagents to perform 96 wells of the kit specific target.
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
phospho-AKT 1/2/3 (Thr308) Capture Antibody Reagent (Part No. IOPAKTBCAR2) - 3 mL
phospho-AKT 1/2/3 (Thr308) Detection Antibody Reagent (Part No. IOPAKTBDAR2) - 3 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

Documents

Manuals & protocols