Overview

  • Product name
    Anti-ERK1 + ERK2 antibody [ERK-7D8]
    See all ERK1 + ERK2 primary antibodies
  • Description
    Mouse monoclonal [ERK-7D8] to ERK1 + ERK2
  • Specificity
    ab54230 reacts strongly with both ERK1 and ERK2.
  • Tested applications
    Suitable for: WB, IP, ELISA, EMSA, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide corresponding to Rat ERK1 + ERK2 aa 324-345.
    Sequence:

    EALAHPYLEQYYDPTDEPVAEE


    Database link: P21708

  • Positive control
    • ICC/IF: U-87 MG (human glioblastoma-astrocytoma epithelial cell line) cells. IHC-P: Human breast carcinoma tissue. Mouse stomach tissue. Human cortex tissue. WB: Human granulosa cell line whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab54230 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 44 kDa.
IP Use at 2-5 µg/mg of lysate.
ELISA Use a concentration of 0.1 - 1 µg/ml.
EMSA Use at an assay dependent concentration.
IHC-P 1/10 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Involved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK1. Phosphorylates EIF4EBP1; required for initiation of translation. Phosphorylates microtubule-associated protein 2 (MAP2). Phosphorylates SPZ1 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4) and ARHGEF2.
    Acts as a transcriptional repressor. Binds to a [GC]AAA[GC] consensus sequence. Repress the expression of interferon gamma-induced genes. Seems to bind to the promoter of CCL5, DMP1, IFIH1, IFITM1, IRF7, IRF9, LAMP3, OAS1, OAS2, OAS3 and STAT1. Transcriptional activity is independent of kinase activity.
  • Sequence similarities
    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • Domain
    The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • Post-translational
    modifications
    Dually phosphorylated on Thr-185 and Tyr-187, which activates the enzyme. Dephosphorylated by PTPRJ at Tyr-187.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • ERK 1 antibody
    • ERK 2 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 2 antibody
    • MAP kinase 1 antibody
    • MAP kinase 2 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
    • Mitogen-activated protein kinase 1 antibody
    • Mitogen-activated protein kinase 2 antibody
    • MK01_HUMAN antibody
    • p38 antibody
    • p40 antibody
    • p41 antibody
    • p42-MAPK antibody
    • PRKM 2 antibody
    see all

Images

  • All lanes : Anti-ERK1 + ERK2 antibody [ERK-7D8] (ab54230) at 1/1000 dilution

    Lane 1 : Human granulosa cell line whole cell lysate
    Lane 2 : Human granulosa cell line whole cell lysate

    Lysates/proteins at 60 µg per lane.

    Secondary
    IRDye® 680-conjugated goat anti-mouse IgG1 (H+L) polyclonal at 1/25000 dilution

    Performed under reducing conditions.

    Predicted band size : 44 kDa
    Observed band size : 44 kDa


    Exposure time : 4 minutes

    This image is courtesy of an Abreview submitted by Francesco Elia Marino

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of ERK1 + ERK2 Antibody (ab54230) was done on 70% confluent log phase U87-MG cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ERK1 + ERK2 Antibody  [ERK-7D8] (ab54230) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488® Rabbit Anti-Mouse IgG Secondary Antibody at a dilution of 1/400 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with DAPI. F-actin (Panel c: red) was stained with Alexa Fluor 594® Phalloidin. Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling ERK1 + ERK2 with ab54230. Staining in the cytoplasm and nucleus (right), compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a ERK1/2 monoclonal antibody (ab54230) diluted in 3% BSA-PBS at a dilution of 1/50 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse stomach tissue labeling ERK1 + ERK2 with ab54230. Staining in the cytoplasm and nucleus (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2 -methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a ERK1/2 monoclonal antibody (ab54230) diluted in 3% BSA-PBS at a dilution of 1/20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cortex tissue labeling ERK1 + ERK2 with ab54230.

References

This product has been referenced in:
  • Chen M  et al. Isoquercetin activates the ERK1/2-Nrf2 pathway and protects against cerebral ischemia-reperfusion injury in vivo and in vitro. Exp Ther Med 13:1353-1359 (2017). WB ; Rat . Read more (PubMed: 28413477) »
  • Ding Y  et al. Effects of 1,25(OH)2 D3 and vitamin D receptor on peripheral CD4(+) /CD8(+) double-positive T lymphocytes in a mouse model of systemic lupus erythematosus. J Cell Mol Med 21:975-985 (2017). WB ; Mouse . Read more (PubMed: 28063200) »

See all 11 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Sample
Human Cell (Human granulosa cell Line)
Specification
Human granulosa cell Line
Permeabilization
Yes - Ethanol and Triton
Fixative
Paraformaldehyde
Username

Dr. Francesco Elia Marino

Verified customer

Submitted Jun 19 2014

Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Non-Denaturing (Native) (12)
Sample
Human Cell lysate - whole cell (Human Granulosa cell line)
Specification
Human Granulosa cell line
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Username

Dr. Francesco Elia Marino

Verified customer

Submitted Jun 02 2014

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

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