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ERK2 Mouse Monoclonal Antibody [A10D2]

Usd: 350

Specification

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Catalog# EM1901-54

ERK2 Mouse Monoclonal Antibody [A10D2]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

ERK2 Mouse Monoclonal Antibody [A10D2]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within human ERK2 aa 200-360.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC

Molecular Weight

Predicted band size: 41 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, A549 cell lysate, A431 cell lysate, HepG2 cell lysate, HEK-293 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, human brain tissue lysate, mouse brain tissue lysate, rat brain tissue lysate, human thyroid tissue, human skin tissue, human breast carcinoma tissue, human pancreas tissue, mouse testis tissue, mouse colon tissue, mouse ovary tissue, K562.

Conjugation

unconjugated

Clone Number

A10D2

RRID

AB_3068899

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

Mitogen-activated protein kinase (MAPK) signaling pathways involve two closely related MAP kinases, known as extracellular-signal-related kinase 1 (ERK 1, p44) and 2 (ERK 2, p42). Growth factors, steroid hormones, G protein-coupled receptor ligands, and neurotransmitters can initiate MAPK signaling pathways. Activation of ERK1 and ERK2 requires phosphorylation by upstream kinases such as MAP kinase kinase (MEK), MEK kinase and Raf-1. ERK1 and ERK2 phosphorylation can occur at specific tyrosine and threonine sites mapping within consensus motifs that include the Threonine-Glutamate-Tyrosine motif. ERK activation leads to dimerization with other ERKs and subsequent localization to the nucleus. Active ERK dimers phosphorylate serine and threonine residues on nuclear proteins and influence a host of responses that include proliferation, differentiation, transcription regulation and development. The human ERK2 gene maps to chromosome 22q11.21 and encodes a 360-amino acid protein.

Background References

1. Wang VY. et. al. Bcl3 Phosphorylation by Akt, Erk2, and IKK Is Required for Its Transcriptional Activity. Mol Cell. 2017 Aug 3;67(3):484-497.e5.

2. Schwebs DJ. et. al. Dictyostelium Erk2 is an atypical MAPK required for chemotaxis. Cell Signal. 2018 Jun;46:154-165.

Sequence Similarity

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-185 and Tyr-187, which activates the enzyme. Undergoes regulatory phosphorylation on additional residues such as Ser-246 and Ser-248 in the kinase insert domain (KID) These phosphorylations, which are probably mediated by more than one kinase, are important for binding of MAPK1/ERK2 to importin-7 (IPO7) and its nuclear translocation. In addition, autophosphorylation of Thr-190 was shown to affect the subcellular localization of MAPK1/ERK2 as well. Ligand-activated ALK induces tyrosine phosphorylation. Dephosphorylated by PTPRJ at Tyr-187. Phosphorylation on Ser-29 by SGK1 results in its activation by enhancing its interaction with MAP2K1/MEK1 and MAP2K2/MEK2. DUSP3 and DUSP6 dephosphorylate specifically MAPK1/ERK2 and MAPK3/ERK1 whereas DUSP9 dephosphorylates a broader range of MAPKs. Dephosphorylated by DUSP1 at Thr-185 and Tyr-187.; ISGylated.

Subcellular Location

Nucleus, spindle, centrosome, cytoplasm, caveola.

UNIPROT

SwissProt: P28482 Human

SwissProt: P63085 Mouse

SwissProt: P63086 Rat

Synonyms

ERK 2 antibody

ERK-2 antibody

ERT1 antibody

Extracellular Signal Regulated Kinase 2 antibody

Extracellular signal-regulated kinase 2 antibody

MAP kinase 1 antibody

MAP kinase 2 antibody

MAP kinase isoform p42 antibody

MAPK 1 antibody

MAPK 2 antibody

Expand

Images

  • Western blot analysis of ERK2 on different lysates with Mouse anti-ERK2 antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: Jurkat cell lysate (20 µg/Lane)<br />Lane 3: A549 cell lysate (20 µg/Lane)<br />Lane 4: A431 cell lysate (20 µg/Lane)<br />Lane 5: HepG2 cell lysate (20 µg/Lane)<br />Lane 6: HEK-293 cell lysate (20 µg/Lane)<br />Lane 7: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 8: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 9: C6 cell lysate (20 µg/Lane)<br />Lane 10: PC-12 cell lysate (20 µg/Lane)<br />Lane 11: Human brain tissue lysate (40 µg/Lane)<br />Lane 12: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 13: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 41 kDa<br />Observed band size: 41 kDa<br /><br />Exposure time: Lane 1-11: 1 minute 30 seconds; Lane 12-13: 15 seconds;<br />ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Alpaca Anti-Mouse IgG - HRP for IP Nano-Secondary Antibody (<a href="/products/NBI02H" style="font-weight: bold;text-decoration: underline;">NBI02H</a>) at 1/5,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ERK2 on different lysates with Mouse anti-ERK2 antibody (EM1901-54) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: Jurkat cell lysate (20 µg/Lane)
    Lane 3: A549 cell lysate (20 µg/Lane)
    Lane 4: A431 cell lysate (20 µg/Lane)
    Lane 5: HepG2 cell lysate (20 µg/Lane)
    Lane 6: HEK-293 cell lysate (20 µg/Lane)
    Lane 7: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 8: RAW264.7 cell lysate (20 µg/Lane)
    Lane 9: C6 cell lysate (20 µg/Lane)
    Lane 10: PC-12 cell lysate (20 µg/Lane)
    Lane 11: Human brain tissue lysate (40 µg/Lane)
    Lane 12: Mouse brain tissue lysate (40 µg/Lane)
    Lane 13: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 41 kDa
    Observed band size: 41 kDa

    Exposure time: Lane 1-11: 1 minute 30 seconds; Lane 12-13: 15 seconds;
    ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-54) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Alpaca Anti-Mouse IgG - HRP for IP Nano-Secondary Antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 1/100) 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 thyroid tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (EM1901-54, 1/100) 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 skin tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 1/100) 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 skin tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (EM1901-54, 1/100) 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 breast carcinoma tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 1/100) 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 breast carcinoma tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (EM1901-54, 1/100) 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 pancreas tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 1/100) 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 pancreas tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (EM1901-54, 1/100) 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 mouse testis tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 1/100) 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 mouse testis tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (EM1901-54, 1/100) 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 mouse colon tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 1/100) 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 mouse colon tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (EM1901-54, 1/100) 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 mouse ovary tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 1/100) 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 mouse ovary tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (EM1901-54, 1/100) 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 ERK2 was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>, 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-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of ERK2 was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-54, 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-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ERK2 on different lysates with Mouse anti-ERK2 antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate (10 µg/Lane)<br />Lane 2: A549-si ERK2 cell lysate (10 µg/Lane)<br /><br />Predicted band size: 41 kDa<br />Observed band size: 41 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1901-54" style="font-weight: bold;text-decoration: underline;">EM1901-54</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of ERK2 on different lysates with Mouse anti-ERK2 antibody (EM1901-54) at 1/2,000 dilution.

    Lane 1: A549-si NT cell lysate (10 µg/Lane)
    Lane 2: A549-si ERK2 cell lysate (10 µg/Lane)

    Predicted band size: 41 kDa
    Observed band size: 41 kDa

    Exposure time: 1 minute; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-54) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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