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  1. Link

    erk1--erk2-antibody-epr17526-ab184699.pdf

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Neuroscience Neurology process Neurodegenerative disease Alzheimer's disease Tangles & Tau
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
  • Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17526] to ERK1 + ERK2
  • Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free HRP PE

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Knockout
Product image
Human MAPK1 (ERK2) knockout HeLa cell line (ab265052)

View more associated products

Overview

  • Product name

    Anti-ERK1 + ERK2 antibody [EPR17526]
    See all ERK1 + ERK2 primary antibodies
  • Description

    Rabbit monoclonal [EPR17526] to ERK1 + ERK2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human ERK1 full length recombinant protein; Human ERK2 full length recombinant protein; A431, Jurkat, HeLa, HepG2, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart, kidney and spleen lysates. ICC/IF: HeLa cells. Flow Cyt (intra): A431 cells. IP. PC-12 whole cell extract.
  • 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.

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: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17526
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Tangles & Tau
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Cell adhesion
    • Other
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-ERK1 + ERK2 antibody [EPR17526] (ab208564)
    • Alexa Fluor® 647 Anti-ERK1 + ERK2 antibody [EPR17526] (ab208881)
    • HRP Anti-ERK1 + ERK2 antibody [EPR17526] (ab209321)
    • Alexa Fluor® 405 Anti-ERK1 + ERK2 antibody [EPR17526] (ab210044)
    • Alexa Fluor® 594 Anti-ERK1 + ERK2 antibody [EPR17526] (ab210460)
    • Alexa Fluor® 555 Anti-ERK1 + ERK2 antibody [EPR17526] (ab210659)
    • PE Anti-ERK1 + ERK2 antibody [EPR17526] (ab212153)
    • Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    • Anti-ERK2 antibody [1B3B9] (ab231085)
    • Anti-ERK2 antibody [1B3B9] - BSA and Azide free (ab269567)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

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

    • Human MAPK1 (ERK2) knockout HeLa cell line (ab265052)
  • KO cell lysates

    • Human MAPK1 (ERK2) knockout HeLa cell lysate (ab257525)
  • Related Products

    • Recombinant Human ERK2 protein (ab43625)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab184699 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
Flow Cyt (Intra)
1/440.

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

IP
1/70.
ICC/IF (1)
1/250.
WB
1/10000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa).
Notes
Flow Cyt (Intra)
1/440.

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

IP
1/70.
ICC/IF
1/250.
WB
1/10000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa).

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.
  • Target information above from: UniProt accession P28482 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5594 Human
    • Entrez Gene: 5595 Human
    • Entrez Gene: 26413 Mouse
    • Entrez Gene: 26417 Mouse
    • Entrez Gene: 116590 Rat
    • Entrez Gene: 50689 Rat
    • Omim: 176948 Human
    • Omim: 601795 Human
    • SwissProt: P27361 Human
    • SwissProt: P28482 Human
    • SwissProt: P63085 Mouse
    • SwissProt: Q63844 Mouse
    • SwissProt: P21708 Rat
    • SwissProt: P63086 Rat
    • Unigene: 431850 Human
    • Unigene: 196581 Mouse
    • Unigene: 34914 Rat
    see all
  • Form

    Mainly expressed in the cytoplasm and only localizes to the nucleus with treatment.
  • Alternative names

    • ERK 1 antibody
    • ERK 2 antibody
    • ERK-1 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 1 antibody
    • Extracellular signal-regulated kinase 2 antibody
    • HS44KDAP antibody
    • HUMKER1A antibody
    • Insulin-stimulated MAP2 kinase antibody
    • MAP kinase 1 antibody
    • MAP kinase 2 antibody
    • MAP kinase 3 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
    • Microtubule-associated protein 2 kinase antibody
    • Mitogen-activated protein kinase 1 antibody
    • Mitogen-activated protein kinase 2 antibody
    • Mitogen-activated protein kinase 3 antibody
    • MK01_HUMAN antibody
    • p38 antibody
    • p40 antibody
    • p41 antibody
    • p41mapk antibody
    • p42-MAPK antibody
    • P42MAPK antibody
    • p44-ERK1 antibody
    • p44-MAPK antibody
    • p44ERK1 antibody
    • p44MAPK antibody
    • PRKM 2 antibody
    • PRKM1 antibody
    • PRKM2 antibody
    • PRKM3 antibody
    • protein tyrosine kinase ERK2 antibody
    see all

Images

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    All lanes : Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    Lane 1 : Mouse brain lysates
    Lane 2 : Mouse heart lysates
    Lane 3 : Mouse kidney lysates
    Lane 4 : Mouse spleen lysates
    Lane 5 : Rat brain lysates
    Lane 6 : Rat heart lysates
    Lane 7 : Rat kidney lysates
    Lane 8 : Rat spleen lysates
    Lane 9 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 10 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 11 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 12 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 44, 42 kDa why is the actual band size different from the predicted?



    44kDa band represents ERK1. 42kDa band represents ERK2.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    All lanes : Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MAPK1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 43, 41 kDa
    Observed band size: 44 kDa why is the actual band size different from the predicted?



    Lanes 1-2: Merged signal (red and green). Green - ab184699 observed at 44 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab184699 Anti-ERK1 + ERK2 antibody [EPR17526] was shown to specifically react with ERK2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265052 (knockout cell lysate ab257525) was used. Wild-type and ERK2 knockout samples were subjected to SDS-PAGE. ab184699 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 Dilution and 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-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)This image is courtesy of an Abreview submitted by Kirk Mcmanus.

    Ab184699 staining ERK1 + ERK2 in HeLa cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. A Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) was used as the secondary antibody.

    See Abreview

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution + Recombinant Human ERK1 protein (ab43623) (ab43623) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 44 kDa why is the actual band size different from the predicted?



    Recombinant full length ERK1 protein (ab43623) contains aa1-379.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ERK1 + ERK2 with ab184699 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing both nuclear and cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab184699 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution + Recombinant Human ERK2 protein (ab43625) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 42 kDa why is the actual band size different from the predicted?



    Recombinant full length ERK2 protein (ab43625) contains aa1-360.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    All lanes : Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/50000 dilution

    Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 4 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysates
    Lane 5 : Human fetal brain lysates
    Lane 6 : Human fetal heart lysates
    Lane 7 : Human fetal kidney lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 42,44 kDa why is the actual band size different from the predicted?



    44kDa band represents ERK1. 42kDa band represents ERK2.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    All lanes : Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    Lane 1 : Human fetal brain lysates
    Lane 2 : Human fetal heart lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 42,44 kDa why is the actual band size different from the predicted?



    44kDa band represents ERK1. 42kDa band represents ERK2.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Intracellular flow cytometric analysis of A431 (Human epidermoid carcinoma) cells labeling ERK1 + ERK2 with ab184699 at 1/440 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody. 

     

     

  • Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    ERK1 + ERK2 were immunoprecipitated from 1mg of PC-12 (Rat adrenal gland pheochromocytoma) whole cell extract with ab184699 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab184699 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: PC-12 whole cell extract. Lane 2: PBS instead of PC-12 whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    44kDa band represents ERK1. 42kDa band represents ERK2.

  • Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (121)

Publishing research using ab184699? Please let us know so that we can cite the reference in this datasheet.

ab184699 has been referenced in 121 publications.

  • Shi L  et al. Spaceflight and simulated microgravity suppresses macrophage development via altered RAS/ERK/NF?B and metabolic pathways. Cell Mol Immunol 18:1489-1502 (2021). PubMed: 31900461
  • He XL  et al. Solid lipid nanoparticles loading with curcumin and dexanabinol to treat major depressive disorder. Neural Regen Res 16:537-542 (2021). PubMed: 32985484
  • Wang X  et al. C-C chemokine receptor 5 signaling contributes to cardiac remodeling and dysfunction under pressure overload. Mol Med Rep 23:N/A (2021). PubMed: 33200795
  • Fei J  et al. Inhibition of SNCG suppresses the proliferation of lung cancer cells induced by high glucose. Mol Med Rep 23:N/A (2021). PubMed: 33313952
  • Hou TY  et al. JNK and p38 Inhibitors Prevent Transforming Growth Factor-ß1-Induced Myofibroblast Transdifferentiation in Human Graves' Orbital Fibroblasts. Int J Mol Sci 22:N/A (2021). PubMed: 33799469
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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Immunocytochemistry/ Immunofluorescence abreview for Anti-ERK1 + ERK2 antibody [EPR17526]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100
Specification
HeLa
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Kirk Mcmanus

Verified customer

Submitted Jul 26 2017

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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