Phospho-ERK1/2 (Thr202/Tyr204) Translocation Assay Kit (Cell-Based) (BN00920)

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BN00920
  • Phospho-ERK1/2 (Thr202/Tyr204) Translocation Assay Kit (Cell-Based) (BN00920)
  • Phospho-ERK1/2 (Thr202/Tyr204) Translocation Assay Kit (Cell-Based) (BN00920)
$756

Description

ELISA Kit Technical ManualMSDS

Phospho-ERK1/2 (Thr202/Tyr204) Translocation Assay Kit (Cell-Based)

Mitogen-activated protein kinases (MAPKs) are proline-directed serine and threonine protein kinases that regulate numerous physiological cell responses including: embryogenesis, cell differentiation, proliferation, migration, apoptosis and death. Extracellular signal-regulated kinases (ERKs) 1 and 2 (ERK1/2), also known as p44 MAPK and p42 MAPK respectively, belong to one of the five major groups of MAPKs. Closely-related ERK1/2 isoforms are uniquely activated by several extracellular signals including growth factors, cytokines, hormones, and neuro-transmitters. Activation of ERK1/2 by the upstream kinases MEK1 and MEK2 occurs via dual phosphorylation on specific threonine (Thr202) and tyrosine (Tyr204) residues on the T*EY* motif. MEK1 and MEK2 are activated through receptors (tyrosine kinases or integrins) via pathways involving adaptor proteins, guanine nucleotide exchange factors, small GTP binding proteins, and MAPKKs. Activated ERK1/2 phosphorylates both, cytosolic (SOS, MNK1/2, RSKs) and nuclear targets. In the nucleus, it affects gene expression and DNA replication by the phosphorylation of MSK 1 and 2 and the transcription factors Elk-1, Sap1, and Sap2. In cultured cells, growth factors or mitogens induce rapid and transient translocation of activated ERK1/2 to nucleus. Different cell lines exhibit various duration, magnitude, and subcellular localization of activated/phosphorylated ERK1/2. The response of the protein may differ even within the same cell line depending on the dose and cell density. Assay Genie's Phospho-ERK1/2 (Thr202/Tyr204) Translocation Assay Kit provides a simple and complete assay in a ready-to-use format to visualize the translocation of activated ERK1/2 between cytoplasmic and nuclear compartments in mammalian cells.

Figures: Tamoxifen-induced translocation of phosphorylated ERK1/2 in MCF-7 cells. MCF-7 cells (1X10 5 cells per well) were grown, fixed and stained according to the included protocol. Figure 1: Cells grown in media supplemented with 10% FBS and treated with a vehicle (A) or 1X Tamoxifen for 20 min (B). Immunofluorescent staining revealed translocation of phosphorylated ERK1/2 from the cytoplasm (A) to nuclei (B). Figure 2: Cells grown in presence of 1% FBS in absence (A) or presence (B) of 1X Tamoxifen for 20 min exhibited translocation of phosphorylated ERK1/2 from nuclei (A) to cytoplasm (B). Bottom panels in Figures 1 and 2 show nuclear staining with DAPI.

Key Information Description

Product SKU

BN00920

Size

50 Assays

Detection Method

Fluorescence microscope capable of measuring EX at 570 nm and equipped with UV filter for DAPI

Species Reactivity

Human, mouse, rat

Applications

Detection of nuclear translocation of phospo-ERK1 and phospo-ERK2 in mammalian cells.

Features and Benefits

  • Detection of subcellular localization of activated ERK1/2 in mammalian cells
  • Screening and characterizing effectors of ERK1/2 kinases
  • Studying of cell signaling, cell division and cell proliferation mechanism

Kit Components

  • Fixative Solution
  • Blocking Buffer
  • Wash Buffer
  • Phospho-ERK1/2 Primary Antibody (100X)
  • Secondary Antibody (100X)
  • Tamoxifen (1000X)
  • DAPI (1000X)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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