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Neuroscience Neurotransmission Intracellular Signaling Kinases
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ERK5 antibody [EP791Y] (ab40809)

  • Datasheet
Reviews (1)Q&A (1)References (7)

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Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Flow Cytometry - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Flow Cytometry - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Immunoprecipitation - Anti-ERK5 antibody [EP791Y] (ab40809)
  • Anti-ERK5 antibody [EP791Y] (ab40809)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP791Y] to ERK5
  • Suitable for: WB, Flow Cyt, IP, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-ERK5 antibody [EP791Y]
    See all ERK5 primary antibodies
  • Description

    Rabbit monoclonal [EP791Y] to ERK5
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ERK5 aa 800-900 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, HAP1, NIH/3T3 and PC-12 cell lysates. Flow Cyt: HeLa and A549 cells. ICC: HeLa cells. IP: HeLa cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP791Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Intracellular Signaling
    • Kinases
    • Neuroscience
    • Neurology process
    • Neural Signal Transduction
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases

Associated products

  • Alternative Versions

    • Anti-ERK5 antibody [EP791Y] - BSA and Azide free (ab232538)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human MAPK7 (ERK5) knockout HeLa cell line (ab265508)
  • KO cell lysates

    • Human MAPK7 (ERK5) knockout HeLa cell lysate (ab258042)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant human ERK5 protein (ab126913)

Applications

Our Abpromise guarantee covers the use of ab40809 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 1/10000. Detects a band of approximately 115 kDa (predicted molecular weight: 89 kDa).

For unpurified use at 1/1000 - 1/5000 dilution. 

Flow Cyt 1/50.

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

For unpurified use at 1/100 dilution. 

IP 1/30.

For unpurified use at 1/50 dilution.

ICC/IF 1/100.

For unpurified use at 1/250 - 1/500 dilution.

Target

  • Function

    Plays a role in various cellular processes such as proliferation, differentiation and cell survival. The upstream activator of MAPK7 is the MAPK kinase MAP2K5. Upon activation, it translocates to the nucleus and phosphorylates various downstream targets including MEF2C. EGF activates MAPK7 through a Ras-independent and MAP2K5-dependent pathway. May have a role in muscle cell differentiation. May be important for endothelial function and maintenance of blood vessel integrity. MAP2K5 and MAPK7 interact specifically with one another and not with MEK1/ERK1 or MEK2/ERK2 pathways.
  • Tissue specificity

    Expressed in many adult tissues. Abundant in heart, placenta, lung, kidney and skeletal muscle. Not detectable in liver.
  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • Domain

    The second proline-rich region may interact with actin targeting the kinase to a specific location in the cell.
    The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • Post-translational
    modifications

    Dually phosphorylated on Thr-219 and Tyr-221, which activates the enzyme (By similarity). Autophosphorylated in vitro on threonine and tyrosine residues when the C-terminal part of the kinase, which could have a regulatory role, is absent.
  • Cellular localization

    Cytoplasm. Nucleus. Translocates to the nucleus upon activation.
  • Target information above from: UniProt accession Q13164 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5598 Human
    • Entrez Gene: 23939 Mouse
    • Entrez Gene: 114509 Rat
    • Omim: 602521 Human
    • SwissProt: Q13164 Human
    • SwissProt: Q9WVS8 Mouse
    • SwissProt: P0C865 Rat
    • Unigene: 150136 Human
    • Unigene: 38172 Mouse
    • Unigene: 144629 Rat
    see all
  • Alternative names

    • Big MAP kinase 1 antibody
    • BMK 1 antibody
    • BMK 1 kinase antibody
    • BMK-1 antibody
    • BMK1 antibody
    • BMK1 Kinase antibody
    • EC 2.7.11.24 antibody
    • ERK 4 antibody
    • ERK 5 antibody
    • ERK-5 antibody
    • ERK4 antibody
    • ERK5 antibody
    • Extracellular signal regulated kinase 5 antibody
    • Extracellular signal-regulated kinase 5 antibody
    • MAP kinase 7 antibody
    • MAPK 7 antibody
    • MAPK7 antibody
    • Mitogen activated protein kinase 7 antibody
    • Mitogen-activated protein kinase 7 antibody
    • MK07_HUMAN antibody
    • OTTHUMP00000065906 antibody
    • OTTHUMP00000065907 antibody
    • PRKM 7 antibody
    • PRKM7 antibody
    • PROTEIN KINASE, MITOGEN-ACTIVATED, 7 antibody
    see all

Images

  • Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
    Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
    All lanes : Anti-ERK5 antibody [EP791Y] (ab40809) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MAPK7 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 89 kDa
    Observed band size: 115 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab40809 observed at 115 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab40809 was shown to react with ERK5 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265508 (knockout cell lysate ab258042) was used. Wild-type HeLa and MAPK7 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40809 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK5 antibody [EP791Y] (ab40809)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK5 antibody [EP791Y] (ab40809)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ERK5 with Purified ab40809 at 1:100 (4.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry - Anti-ERK5 antibody [EP791Y] (ab40809)
    Flow Cytometry - Anti-ERK5 antibody [EP791Y] (ab40809)

    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ERK5 with purified ab40809 at 1:50 dilution (10 ug/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
    Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
    All lanes : Anti-ERK5 antibody [EP791Y] (ab40809) at 1/2000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates with 5% NFDM/TBST
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates with 5% NFDM/TBST

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 89 kDa
    Observed band size: 115 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
    Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: MAPK7 (ERK5) knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab40809 observed at 88 kDa. Red - loading control, ab9484, observed at 37 kDa.

    Unpurified ab40809 was shown to specifically react with ERK5 in wild-type HAP1 cells as signal was lost in MAPK7 (ERK5) knockout cells. Wild-type and MAPK7 (ERK5) knockout samples were subjected to SDS-PAGE. Unpurified ab40809 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
    Western blot - Anti-ERK5 antibody [EP791Y] (ab40809)
    Anti-ERK5 antibody [EP791Y] (ab40809) at 1/5000 dilution (unpurified) + Hela cell lysate at 10 µg

    Predicted band size: 89 kDa
    Observed band size: 115 kDa why is the actual band size different from the predicted?



    The predicted weight of 89 kDa is for the precursor version of human ERK5 protein. However, ab40809 detects endogenous levels of total Erk5 protein which appears around 115 kDa in SDS PAGE
  • Flow Cytometry - Anti-ERK5 antibody [EP791Y] (ab40809)
    Flow Cytometry - Anti-ERK5 antibody [EP791Y] (ab40809)

    Overlay histogram showing A549 cells stained with unpurified ab40809 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (ab40809, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/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.

  • Immunoprecipitation - Anti-ERK5 antibody [EP791Y] (ab40809)
    Immunoprecipitation - Anti-ERK5 antibody [EP791Y] (ab40809)

    ab40809 (purified) at 1:30 dilution (2ug) immunoprecipitating ERK5 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab40809 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40809 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Anti-ERK5 antibody [EP791Y] (ab40809)
    Anti-ERK5 antibody [EP791Y] (ab40809)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (7)

    Publishing research using ab40809? Please let us know so that we can cite the reference in this datasheet.

    ab40809 has been referenced in 7 publications.

    • Chen R  et al. Cx43 and AKAP95 regulate G1/S conversion by competitively binding to cyclin E1/E2 in lung cancer cells. Thorac Cancer 11:1594-1602 (2020). PubMed: 32338437
    • Song B  et al. LncRNA ENST00000539653 acts as an oncogenic factor via MAPK signalling in papillary thyroid cancer. BMC Cancer 19:297 (2019). PubMed: 30940124
    • Li L  et al. Thymic Stromal Lymphopoietin Promotes Fibrosis and Activates Mitogen-Activated Protein Kinases in MRC-5 Cells. Med Sci Monit 22:2357-62 (2016). WB ; Human . PubMed: 27385084
    • Xiong Y  et al. Phosphorylation of BMK1 induces prostatic carcinoma cell proliferation by promoting entry into the S phase of the cell cycle. Oncol Lett 11:99-104 (2016). PubMed: 26870175
    • Kuss M  et al. Interferon-? induces leucine-rich repeat kinase LRRK2 via extracellular signal-regulated kinase ERK5 in macrophages. J Neurochem 129:980-7 (2014). PubMed: 24479685
    • Yoshizuka N  et al. A novel function of p38-regulated/activated kinase in endothelial cell migration and tumor angiogenesis. Mol Cell Biol 32:606-18 (2012). PubMed: 22124154
    • Takaoka Y  et al. Forced expression of miR-143 represses ERK5/c-Myc and p68/p72 signaling in concert with miR-145 in gut tumors of Apc(Min) mice. PLoS One 7:e42137 (2012). PubMed: 22876303

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-ERK5 antibody [EP791Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (melanoma cell line)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Specification
    melanoma cell line
    Blocking step
    (agent) for 50 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Dr. 令 中野

    Verified customer

    Submitted Jun 20 2019

    Question

    Thanks, but is one of them, suitable after formaldehyde fixing before ChIP? because i see a note referred to the monoclonal Ab that is not clear:

    Application notes
    Is unsuitable for or IHC

    Read More

    Abcam community

    Verified customer

    Asked on Mar 09 2012

    Answer

    Following up my previous message, the lab confirmed that ab40809 was negative for IHC on formalin -fixed tissue. However, it was positive for ICC and flow cytometry on PFA-fixed samples. These application use relatively short fixation times compared to IHC samples. It is possible that the relatively shortcross-linking step for ChIP will also allow a positive result, but we have no data.

    Read More

    Abcam Scientific Support

    Answered on Mar 09 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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