ERK1/ERK2 (Total/Phospho) Multispecies InstantOne™ ELISA Kit

Catalog number: 85-86013-11

Invitrogen™  Related applications: Protein Assays and Analysis

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Description

The InstantOne™ ELISA is specifically engineered for accurate measurement of total and phosphorylated human, mouse, and hamster ERK 1/2 in cell lysates. Detection of rat ERK 1/2 is 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. In the classical MAPK pathway, ligand binding induces the activation of an RTK that initiates a signaling cascade that results in activation of the GTPase Ras and the Ser/Thr kinase Raf (MAPKKK). Raf then binds to and activates MEK (MAPKK) via phosphorylation. 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. Both phosphorylation sites are required for ERK activation and no known mutations exist that cause constitutive ERK1/2 activation. As such, detection of ERK1/2 phosphorylation is frequently used to assess its activation, as well as that of MEK, a popular drug target. Once activated, ERK phosphorylates a variety of proteins that regulate cellular processes such as cell division, proliferation, survival, differentiation, apoptosis, motility, and metabolism. Due to its established critical role in these processes, the MAPK signaling pathway is a frequent target for oncology-related drug development.

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: 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: 5594, 5595
Gene Name: mitogen-activated protein kinase 1, mitogen-activated protein kinase 3
Gene Symbol: 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
Target (Kit Named): ERK1/ERK2
Target Species (Named): Multispecies
Target Species (Validated): Hamster, Human, Mouse
Target Sub-Specificity: Phospho, Total
UniProt ID: P27361, P28482

Contents & storage

The ERK 1/2 (Total/Phospho) InstantOne ELISA contains sufficient reagents to detect a total of 96 wells (48 wells of total ERK 1/2 and 48 wells of phospho-ERK 1/2)
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
ERK 1/2 (Total) Capture Antibody Reagent (Part No. IOERKCAR1M) – 1 x 0.75 mL
phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) Capture Antibody Reagent (Part No. IOPERKCAR1M)–3 x 0.75 mL
ERK 1/2 (Total) Detection Antibody Reagent (Part No. IOERKDAR1M) – 1 x 0.75 mL
phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) Detection Antibody Reagent (Part No. IOPERKDAR1M)–3 x 0.75 mL
Positive Control Cell Lysate (lyophilized) (Part No. IOERKPC1) - 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