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LanthaScreen™ ERK2 A375 Cell Line

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.

ERK1/ERK2 (Phospho) [pT202/pY204] Multispecies ELISA Kit (Invitrogen™)

ERK1/ERK2 (Phospho) [pT202/pY204] ELISA Kit for ELISA

LanthaScreen™ Tb-anti-ERK2 [pThr185/pTyr187] Antibody

A part of the MAPK Pathway, ERK2 is phosphorylated at Thr185 and Tyr187 by the mitogen-activated protein kinase kinases 2 (MAP2K2) .
For use in combination with the LanthaScreen® ERK2 A375 cell line (K1798) to evaluate inhibitors in medium to high throughput screening applications.

ERK1/ERK2 Antibody (6HCLC), ABfinity™ Rabbit Oligoclonal (Invitrogen™)

ERK1/ERK2 Oligoclonal Antibody for Western Blot, ChIP

Phospho-ERK1/ERK2 (Thr185, Tyr187) Antibody (15H10L7), ABfinity™ Rabbit Monoclonal (Invitrogen™)

Phospho-ERK1/ERK2 (Thr185, Tyr187) Monoclonal Antibody for Western Blot, IF, ICC, IHC (P), ChIP, ELISA

ERK1/ERK2 (Phospho) [pT202/pY204]/[pT185/pY187] Multispecies InstantOne™ ELISA Kit (Invitrogen™)

ERK1/ERK2 (Phospho) [pT202/pY204]/[pT185/pY187] Multispecies InstantOne™ ELISA Kit for ELISA

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

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

ERK1/ERK2 Antibody (3HCLC), ABfinity™ Rabbit Oligoclonal (Invitrogen™)

ERK1/ERK2 Oligoclonal Antibody for Western Blot, ChIP

MAPK1 (ERK2) Recombinant Human Protein

MAPK1 (ERK2) is a serine⁄threonine kinase that acts as an important activator of p90 RSK, MSK, ELK1, and Stat3, and is involved in the MAPK pathway.

ERK1/ERK2 Multispecies In-Cell ELISA Kit, Colorimetric (Invitrogen™)

ERK1/ERK2 Multispecies In-Cell ELISA Kit, Colorimetric for ELISA

ERK1/ERK2 Polyclonal Antibody (Invitrogen™)

ERK1/ERK2 Polyclonal Antibody for Western Blot, IF, ICC, IHC (P)

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

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

Phospho-ERK1/ERK2 (Thr185, Tyr187) Polyclonal Antibody, Alexa Fluor 488 (Invitrogen™)

Phospho-ERK1/ERK2 (Thr185, Tyr187) Polyclonal Antibody for IF, ICC

LanthaScreen™ Tb-anti-ERK2 [pThr185/pTyr187] Antibody Kit

To facilitate in-house kinase assay configuration and screening, we have developed a selection of Tb-labeled phospho-specific antibodies, which may be accessed in small scale and screening quantities.

ERK2 Monoclonal Antibody (6F8) (Invitrogen™)

ERK2 Monoclonal Antibody for Western Blot, IF, ICC, IHC (P), Flow, IP, ChIP