PRODUCT CODE: ET1603-22

Recombinant Phospho Erk1 (T202)+Erk2 (T185) Monoclonal Antibody (ET1603-22)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

ICC staining of Phospho-Erk1(T202)+Erk2(T185) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-Erk1(T202)+Erk2(T185) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-Erk1(T202)+Erk2(T185) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-Erk1(T202)+Erk2(T185) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-Phospho-Erk1(T202)+Erk2(T185) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-22, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-Erk1(T202)+Erk2(T185) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-22, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of Phospho-Erk1(T202)+Erk2(T185) was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1603-22, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ICC staining of Phospho-Erk1(T202)+Erk2(T185) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho Erk1 (T202)+Erk2 (T185) Monoclonal Antibody (ET1603-22)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding thr185 of human erk2.

Host

Rabbit

Modification

Phospho

Modification Site

Erk1(T202)+Erk2(T185)

Positive Control

A549, NIH/3T3, MCF-7, human lung carcinoma tissue, human kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SZ2-4

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

42/44 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Phospho-Erk1(T202)+Erk2(T185)

SYNONYMS

ERK 1 antibody; ERK 2 antibody; ERK-1 antibody; ERK-2 antibody; ERK1 antibody; erk1/2 antibody; ERK2 antibody; ERT1 antibody; ERT2 antibody; Extracellular signal regulated kinase 1 antibody; Extracellular signal-regulated kinase 1 antibody; Extracellular signal-regulated kinase 2 antibody; HS44KDAP antibody; HUMKER1A antibody; Insulin-stimulated MAP2 kinase antibody; MAP kinase 1 antibody; MAP kinase 2 antibody; MAP kinase 3 antibody; MAP kinase isoform p42 antibody; MAP kinase isoform p44 antibody; MAPK 1 antibody; MAPK 2 antibody; MAPK 3 antibody; Mapk1 antibody; MAPK2 antibody; MAPK3 antibody; Microtubule-associated protein 2 kinase antibody; Mitogen-activated protein kinase 1 antibody; Mitogen-activated protein kinase 2 antibody; Mitogen-activated protein kinase 3 antibody; MK01_HUMAN antibody; p38 antibody; p40 antibody; p41 antibody; p41mapk antibody; p42-MAPK antibody; P42MAPK antibody; p44-ERK1 antibody; p44-MAPK antibody; p44ERK1 antibody; p44MAPK antibody; PRKM 2 antibody; PRKM1 antibody; PRKM2 antibody; PRKM3 antibody; protein tyrosine kinase ERK2 antibody

SEQUENCE SIMILARITIES

Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated upon KIT and FLT3 signaling (By similarity). Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Ligand-activated ALK induces tyrosine phosphorylation. Dephosphorylated by PTPRJ at Tyr-204.

SUBCELLULAR LOCATION

Cytoplasm, Nucleus.

FUNCTION

Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines, and research investigators consider it an important target in the diagnosis and treatment of cancer. Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway. MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK and the transcription factor Elk-1. p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs, along with MEK inhibitors, such as U0126 and PD98059.