AKT/ERK/p70 S6K Activation Multispecies InstantOne™ ELISA Kit

Catalog number: 85-86018-11

Invitrogen™  Related applications: Protein Assays and Analysis

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Description

The InstantOne™ ELISA is specifically engineered for accurate measurement of phosphorylated ERK 1/2 (human, mouse, hamster), AKT 1/2/3 (human, mouse, hamster), and p70 S6K (human, mouse) in cell lysates. Detection of these phosphorylated molecules in rat 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.

Extracellular signal-regulated kinase (ERK) is the founding member and a key component of the classical Mitogen-Activated Protein Kinase (MAPK) pathway. MEK activates ERK 1/2 by phosphorylation of its TxY motifs in response to a variety of stimuli, including growth factors and cytokines.

AKT, which exists as multiple isoforms, is one of the principle kinases activated by phosphoinositide 3-kinase (PI3K). 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.

p70 S6 Kinase (p70S6K) is a member of the ribosomal S6 kinase family of Ser/Thr kinases. p70 S6K activity is controlled by multiple signaling pathways, including the MAPK, phosphoinositide-3 kinase (PI3K), and mTOR pathways. This regulation results in the phosphorylation of multiple sites of p70 S6K, including Thr389 within the linker domain, which is required for full activation. Activated p70 S6K phosphorylates several residues on the S6 ribosomal protein, thereby leading to increased protein synthesis.

This kit has been developed to recognize the following phosphorylation sites:

ERK 1/2/3 (Thr202/Tyr204, Thr185/Tyr187)
AKT 1/2/3 (Ser473)
p70 S6K (Thr389)

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, ERK, ERK-2, ERK2, ERT1, MAPK2, P42MAPK, PRKM1, PRKM2, p38, p40, p41, p41mapk, p42-MAPK, ERK-1, ERK1, ERT2, HS44KDAP, HUMKER1A, P44ERK1, P44MAPK, PRKM3, p44-ERK1, p44-MAPK, PS6K, S6K, S6K-beta-1, S6K1, STK14A, p70 S6KA, p70(S6K)-alpha, p70-S6K, p70-alpha
Gene ID: 207, 5594, 5595, 6198
Gene Name: AKT serine/threonine kinase 1, mitogen-activated protein kinase 1, mitogen-activated protein kinase 3, ribosomal protein S6 kinase B1
Gene Symbol: AKT1, MAPK1, MAPK3, RPS6KB1
Immunoassay Kit Format: Coated ELISA Kit
Label or Dye: HRP (Horseradish Peroxidase)
Product Line: InstantOne™
Product Size: 96 tests
Protein Name: Mitogen-activated protein kinase 1, Mitogen-activated protein kinase 3, RAC-alpha serine/threonine-protein kinase, Ribosomal protein S6 kinase beta-1
Sample Type (Specific): Cell Extracts
Target (Kit Named): AKT/ERK/p70 S6K Activation
Target Species (Named): Multispecies
Target Species (Validated): Human, Mouse
UniProt ID: P23443, P27361, P28482, P31749

Contents & storage

The ERK/AKT/ p70 S6K Activation InstantOne ELISA contains sufficient reagents to detect a total of 96 wells (32 wells for each of the three phosphorylated targets)
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 (Ser437) Capture Antibody Reagent (IOPAKTACAR1T) - 1 x 1.0 mL
phospho-AKT 1/2/3 (Ser437) Detection Antibody Reagent (IOPAKTADAR1T)- 1 x 1.0 mL
phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) Capture Antibody Reagent (IOPERKCAR1T) - 1 x 1.0 mL
phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) Detection Antibody Reagent (IOPERKDAR1T) - 1 x 1.0 mL
phospho-p70 S6K (Thr389) Capture Antibody Reagent (IOPS6KCAR1T) - 1 x 1.0 mL
phospho-p70 S6K (Thr389) Detection Antibody Reagent (IOPS6KDAR1T) - 1 x 1.0 mL
Positive Control Cell Lysate lyophilized (IOMEAPPC1)- 0.25 mL
Detection Reagent (TMB Substrate) (Part No. IODR1) - 10 mL
Stop Solution (Part No. IOSS1) - 10 mL
Plate Seal - 1 ea

Documents

Manuals & protocols