Overview

  • Product name
    Anti-ERK1 + ERK2 antibody [ERK-7D8]
    See all ERK1 + ERK2 primary antibodies
  • Description
    Mouse monoclonal [ERK-7D8] to ERK1 + ERK2
  • Host species
    Mouse
  • 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: Recombinant human ERK1 protein (ab116536), 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

    All lanes : Human granulosa cell line whole cell lysate

    Lysates/proteins at 60 µg per lane.

    Secondary
    All lanes : 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

    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 X  et al. Anti-Semaphorin-7A single chain antibody demonstrates beneficial effects on pulmonary inflammation during acute lung injury. Exp Ther Med 15:2356-2364 (2018). Read more (PubMed: 29456642) »
  • Wang Y  et al. Adipokine RBP4 drives ovarian cancer cell migration. J Ovarian Res 11:29 (2018). Read more (PubMed: 29642915) »
See all 27 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

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

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

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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