AKT/ERK Activation Multispecies InstantOne™ ELISA Kit

Catalog number: 85-86014-11

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 ERK 1/2 and AKT 1/2/3 in cell lysates. Detection of rat ERK 1/2 and AKT 1/2/3 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. ERK1 (p44, MAPK3) and ERK2 (p42, MAPK2) are both activated via the MAPK/ERK pathway, which is downstream of various Receptor Tyrosine Kinases (RTKs) or GPCRs. MEK, a Ser/Thr and Tyr kinase, activates ERK by phosphorylation of its TxY motifs, namely Thr202/Tyr204 and Thr185/Tyr187 of ERK1 and ERK2, respectively. Once activated, ERK phosphorylates a variety of proteins that regulate cellular processes such as cell division, proliferation, survival, differentiation, apoptosis, motility, and metabolism.

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.

This kit has been developed to recognize the following phosphorylation sites:
ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187)
AKT 1/2/3 (Ser473)

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
Gene ID: 207, 5594, 5595
Gene Name: AKT serine/threonine kinase 1, mitogen-activated protein kinase 1, mitogen-activated protein kinase 3
Gene Symbol: AKT1, MAPK1, MAPK3
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
Sample Type (Specific): Cell Extracts
Target (Kit Named): AKT/ERK Activation
Target Species (Named): Multispecies
Target Species (Validated): Hamster, Human, Mouse
UniProt ID: P27361, P28482, P31749

Contents & storage

The AKT/ERK Activation InstantOne ELISA contains sufficient reagents to detect a total of 96 wells (48 wells of phopsho-AKT and 48 wells of phospho-ERK)
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 (IOPAKTACAR1M) - 2 x 0.75 mL
phospho-AKT 1/2/3 (Ser437) Detection Antibody Reagent (IOPAKTADAR1M)- 2 x 0.75 mL
phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) Capture Antibody Reagent (IOPERKCAR1M) - 2 x 0.75 mL
phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) Detection Antibody Reagent (IOPERKDAR1M)- 2 x 0.75 mL
Positive Control Cell Lysate lyophilized - 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