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Thermo Scientific™

LanthaScreen™ ERK2 A375 Cell Line

Catalog number: K1798
Thermo Scientific™

LanthaScreen™ ERK2 A375 Cell Line

Catalog number: K1798
Catalog Number
K1798
Unit Size
1 vial
Price (USD)
8,140.00
Your Price:
Availability
***
Quantity
Catalog NumberUnit SizePrice (USD)AvailabilityQuantity
K17981 vial
8,140.00
Your Price:
***
Product Overview
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The Map Kinase (MapK) signal transduction cascade is activated by growth factors such as EGF, PDGF, and HGF. Binding of these factors to their respective cell surface receptors results in the initiation of receptor tyrosine kinase activity, which leads to the sequential phospho-activation of downstream kinases such as Ras, Raf, MEK, and Erk1⁄2. Activated MEK phosphorylates Erk2 (Mapk1) proteins at a specific Thr-Tyr motif (Thr⁄Tyr 185⁄187). A number of constitutive active kinase mutants within the MAP kinase pathway have been implicated in oncogenesis. One of these mutants is BRAF(V600E), which is the most predominant oncogenic BRAF mutant. The human melanoma cell line A375 endogenously expresses BRAF(V600E), which leads to the constitutive activation of the MAP kinase pathway and phosphorylation of Erk2 in the absence of ligands. LanthaScreenTM Erk2 A375 constitutively expresses GFP-Erk2 under control of a CMV promotor. Using this cell line, a homogenous immuno-assay has been developed in which the phosphorylation state of GFP-Erk2 is detected in cell lysates using a terbium-labeled anti-pTpY-185⁄187-Erk2 antibody, in a time-resolved FRET (TR-FRET) readout. This cell line can be used to evaluate compound activity against BRAF(V600E). GFP-Erk2 lentivirus was transduced into A375 cells, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. Using the lytic TR-FRET immuno-assay, this cell line is validated for EC50 and Z’ under optimized conditions using EGF as a ligand for GFP-Erk2 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and cell lysis⁄equilibration time. Additional information using alternate stimuli and small molecule inhibitor is also provided.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Antagonists
Raf Inhibitor I
Assay Entry
Cell-based terbium phospho-specific activity (stable)
Cell State
Dividing Cells
Detection Method
Fluorescent
For Use With (Application)
TR-FRET
Gene Symbol
ERK2
Primary Agonist
Constitutive
Product Line
LanthaScreen™
Readout
End Point
Target Entry
ERK2
Cell Line
A375
System Type
LanthaScreen™
Druggable Target
Signaling Pathway

Contents & Storage

LanthaScreen™ ERK2 A375 cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt, or thaw for immediate use.

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