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  1. Link

    erk1--erk2-antibody-epr17526-bsa-and-azide-free-ab218017.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

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  • Certificate of Compliance
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Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
  • Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

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 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), ICC/IF, IP, 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 HRP PE Unconjugated

You may also be interested in

Conjugation
Product image
APC Conjugation Kit - Lightning-Link® (ab201807)
Knockout
Product image
Human MAPK1 (ERK2) knockout HeLa cell line (ab265052)
Protein
Product image
Recombinant Human HIF-1 alpha protein (ab48734)

View more associated products

Overview

  • Product name

    Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free
    See all ERK1 + ERK2 primary antibodies
  • Description

    Rabbit monoclonal [EPR17526] to ERK1 + ERK2 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, IP, 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: HeLa cell lysate. ICC/IF: HeLa cells. Flow Cyt (intra): A431 cells. IP. PC-12 whole cell extract.
  • General notes

    ab218017 is the carrier-free version of ab184699.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • 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

    • Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)
    • 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)
  • 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 ab218017 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)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa).
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. 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] - BSA and Azide free (ab218017)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab184699).

    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] - BSA and Azide free (ab218017)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699).

  • Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699).

  • Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Clone EPR17526 (ab218017) has been successfully conjugated by Abcam. This image was generated using Anti-ERK1 + ERK2 antibody [EPR17526] (PE). Please refer to ab212153 for protocol details. 

    Overlay histogram showing A431 cells stained with ab212153 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol at -20°C for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab212153, 1/2500 dilution) for 30 min at 22°C.Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter. 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699).

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Clone EPR17526 (ab218017) has been successfully conjugated by Abcam. This image was generated using Anti-ERK1 + ERK2 antibody [EPR17526] (Alexa Fluor® 647). Please refer to ab208881 for protocol details.

    ab208881 staining ERK1 + ERK2 in A431 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab208881 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Clone EPR17526 (ab218017) has been successfully conjugated by Abcam. This image was generated using Anti-ERK1 + ERK2 antibody [EPR17526] (Alexa Fluor® 488). Please refer to ab208564 for protocol details.

    ab208564 staining ERK1 + ERK2 in A431 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab208564 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in A431 cells fixed with 4% formaldehyde (10 min).

     

  • Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    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. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699). 

     

     

  • Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

Protocols

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

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

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

References (2)

Publishing research using ab218017? Please let us know so that we can cite the reference in this datasheet.

ab218017 has been referenced in 2 publications.

  • Guo J  et al. Berberine Protects Human Umbilical Vein Endothelial Cells against LPS-Induced Apoptosis by Blocking JNK-Mediated Signaling. Evid Based Complement Alternat Med 2016:6983956 (2016). WB ; Human . PubMed: 27478481
  • Xu Y  et al. Mechanisms of PDGF siRNA-mediated inhibition of bone cancer pain in the spinal cord. Sci Rep 6:27512 (2016). WB ; Rat . PubMed: 27282805

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