Overview

  • Product nameAnti-ERK1 antibody [Y72]
    See all ERK1 primary antibodies
  • Description
    Rabbit monoclonal [Y72] to ERK1
  • Specificityab32537 recognises ERK1. The antibody does not cross-react with other MAP kinases.
  • Tested applicationsWB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Recombinant Fragment
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human ERK1 aa 1-100 (N terminal).
    Database link: P27361

  • Epitopeab32537 reacts with an epitope located in the N terminal region of ERK1.
  • Positive control
    • WB: HeLa, HEK293, Jurkat and RAW264.7 whole cell lysates and ERK1 recombinant protein. IHC: Human lung carcinoma, human cervix carcinoma and human tonsil tissues. ICC/IF: Jurkat cells. Flow Cyt: HeLa and Jurkat cells. IP: Jurkat cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    A trial size is available to purchase for this antibody.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative versions available:

    Anti-ERK1 antibody (Alexa Fluor® 488) [Y72] (ab190200)

    Anti-ERK1 antibody (HRP) [Y72] (ab190578)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

Properties

Applications

Our Abpromise guarantee covers the use of ab32537 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 1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 43 kDa).
IP 1/20 - 1/40.
Flow Cyt 1/30 - 1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/100.

Target

  • FunctionInvolved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK-1. Phosphorylates EIF4EBP1; required for initiation of translation. Phosphorylates microtubule-associated protein 2 (MAP2). Phosphorylates SPZ1 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4).
  • Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • DomainThe TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • Post-translational
    modifications
    Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Dephosphorylated by PTPRJ at Tyr-204.
  • Information by UniProt
  • Database links
  • Alternative names
    • ERK 1 antibody
    • ERK antibody
    • ERK-1 antibody
    • ERK1 antibody
    • ERT 2 antibody
    • ERT2 antibody
    • Extracellular Signal Regulated Kinase 1 antibody
    • Extracellular signal related kinase 1 antibody
    • Extracellular signal-regulated kinase 1 antibody
    • HGNC6877 antibody
    • HS44KDAP antibody
    • HUMKER1A antibody
    • Insulin Stimulated MAP2 Kinase antibody
    • Insulin-stimulated MAP2 kinase antibody
    • MAP kinase 1 antibody
    • MAP kinase 3 antibody
    • MAP Kinase antibody
    • MAP kinase isoform p44 antibody
    • MAPK 1 antibody
    • MAPK 3 antibody
    • MAPK antibody
    • MAPK1 antibody
    • Mapk3 antibody
    • MGC20180 antibody
    • Microtubule Associated Protein 2 Kinase antibody
    • Microtubule-associated protein 2 kinase antibody
    • Mitogen Activated Protein Kinase 3 antibody
    • Mitogen-activated protein kinase 1 antibody
    • Mitogen-activated protein kinase 3 antibody
    • MK03_HUMAN antibody
    • OTTHUMP00000174538 antibody
    • OTTHUMP00000174541 antibody
    • p44 ERK1 antibody
    • p44 MAPK antibody
    • p44-ERK1 antibody
    • p44-MAPK antibody
    • P44ERK1 antibody
    • P44MAPK antibody
    • PRKM 3 antibody
    • PRKM3 antibody
    • Protein Kinase Mitogen Activated 3 antibody
    see all

Anti-ERK1 antibody [Y72] images

  • All lanes : Anti-ERK1 antibody [Y72] (ab32537) at 1/10000 dilution (purified)

    Lane 1 : 293T whole cell lysate
    Lane 2 : HeLa whole cell lysate
    Lane 3 : Jurkat whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 43 kDa
    Observed band size : 44 kDa (why is the actual band size different from the predicted?)
    Blocking and dilution buffer: 5% NFDM/TBST
  • Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution (purified) + RAW264.7 whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Predicted band size : 43 kDa
    Observed band size : 44 kDa (why is the actual band size different from the predicted?)
    Blocking and dilution buffer: 5% NFDM/TBST


  • Predicted band size : 43 kDa

    Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Green signal from target - ab32537 observed at 42 kDa
    Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
    Lanes 5 and 6: Merged (red and green) signal
    ab32537 was shown to specifically react with ERK1 when ERK1 knockout samples were used. Wild-type and ERK1 knockout samples were subjected to SDS-PAGE. ab32537 and ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : Recombinant Human ERK1 protein (ab43623)
    Lane 4 : Recombinant Human ERK2 protein (ab43625)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

    Predicted band size : 43 kDa
    Observed band size : 44 kDa (why is the actual band size different from the predicted?)

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab32537 overnight at 4°C. Antibody binding was detected using a goat anti-rabbit Alexa Fluor® 790) ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution (unpurified) + Jurkat cell lysate

    Predicted band size : 43 kDa
    Observed band size : 43 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling ERK1 with purified ab32537 at a dilution of 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • IHC image of ab32537 staining ERK1 in Human tonsil formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32537, 2μg/ml dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling ERK1 with unpurified ab32537.

  • Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling ERK1 with purified ab32537 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Unpurified ab32537 staining ERK1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081, a goat anti rabbit Alexa Fluor® 488 (1/200) was used as the secondary antibody. Counterstained with DAPI. Cytoplasmic and nuclear staining shown.

    See Abreview

  • Flow Cytometry analysis of Jurkat cells labelling ERK1 with purified ab32537 at a dilution of 1/30 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Overlay histogram showing HeLa cells stained with ab32537 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32537, 1/11312) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween 20 for 20 min used under the same conditions.

  • Unpurified ab32537 staining ERK1 (green) in HEK293 cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with 70% methanol. The sample was incubated with the primary antibody (1/40 in PBS + 0.2% BSA + 0.1% sodium azide) for 1 hour at 22°C. A phycoerythrin-conjugated goat anti-rabbit IgG (1/100) was used as the secondary antibody.
    Gating Strategy: Live Cells. Purple plot represents isotype control.

    See Abreview

  • ab32537 (purified) at a dilution of 1/20 immunoprecipitating ERK1 in Jurkat whole cell lysate.

    Lane 1 (input): Jurkat whole cell lysate (10µg)

    Lane 2 (+): ab32537 + Jurkat whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32537 in Jurkat whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

References for Anti-ERK1 antibody [Y72] (ab32537)

This product has been referenced in:
  • Tor YS  et al. Induction of Apoptosis in MCF-7 Cells via Oxidative Stress Generation, Mitochondria-Dependent and Caspase-Independent Pathway by Ethyl Acetate Extract of Dillenia suffruticosa and Its Chemical Profile. PLoS One 10:e0127441 (2015). WB ; Human . Read more (PubMed: 26047480) »
  • Dakhova O  et al. Global gene expression analysis of reactive stroma in prostate cancer. Clin Cancer Res 15:3979-89 (2009). IHC ; Human . Read more (PubMed: 19509179) »

See all 2 Publications for this product

Product Wall

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - 70% methanol
Sample Human Cell (HEK293)
Specification HEK293
Gating Strategy live cells gated
Preparation Cell harvesting/tissue preparation method: Cell dissociation buffer
Sample buffer: Enzyme free
Username

Abcam user community

Verified customer

Submitted Aug 04 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Sample Human Cell (HEK293)
Specification HEK293
Permeabilization Yes - 0.1% Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 04 2014

Application Western blot
Loading amount 50000 cells
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Aug 04 2014

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton X100 in PBS
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

Submitted Mar 27 2014

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