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    erk2-antibody-e460-ab32081.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ERK2 antibody [E460] (ab32081)

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  • SDS
Reviews (2)Q&A (1)References (31)

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Western blot - Anti-ERK2 antibody [E460] (ab32081)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK2 antibody [E460] (ab32081)
  • Western blot - Anti-ERK2 antibody [E460] (ab32081)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK2 antibody [E460] (ab32081)
  • Western blot - Anti-ERK2 antibody [E460] (ab32081)
  • Flow Cytometry (Intracellular) - Anti-ERK2 antibody [E460] (ab32081)
  • Western blot - Anti-ERK2 antibody [E460] (ab32081)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK2 antibody [E460] (ab32081)
  • Anti-ERK2 antibody [E460] (ab32081)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E460] to ERK2
  • Suitable for: IHC-P, Flow Cyt (Intra), WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free HRP

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Human MAPK1 (ERK2) knockout HeLa cell line (ab265052)

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Overview

  • Product name

    Anti-ERK2 antibody [E460]
    See all ERK2 primary antibodies
  • Description

    Rabbit monoclonal [E460] to ERK2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt (Intra), WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Recombinant fragment
  • Immunogen

    Synthetic peptide within Human ERK2 aa 300 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P28482

  • Epitope

    ab32081 reacts with an epitope located in the C terminal region of ERK2.
  • Positive control

    • WB: A431 whole cell lysate (ab7909), HeLa, HEK293, MES and PC12 whole cell lysates and ERK2 recombinant protein. IHC-P: Rat pancreas tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
  • 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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E460
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Intracellular Signaling
    • Kinases
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-ERK2 antibody [E460] (ab190581)
    • HRP Anti-ERK2 antibody [E460] (ab205819)
    • Alexa Fluor® 555 Anti-ERK2 antibody [E460] (ab213333)
    • Anti-ERK2 antibody [E460] - BSA and Azide free (ab214170)
    • 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)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • 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)
  • KO cell pellets

    • Human MAPK1 (ERK2) knockout HeLa cell pellet (ab278969)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human ERK2 protein (ab95379)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32081 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
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.

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

WB (1)
1/1000 - 1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 41 kDa).

Can be blocked with ab205612

ICC/IF
1/400.
Notes
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.

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

WB
1/1000 - 1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 41 kDa).

Can be blocked with ab205612

ICC/IF
1/400.

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: 26413 Mouse
    • Entrez Gene: 116590 Rat
    • Omim: 176948 Human
    • SwissProt: P28482 Human
    • SwissProt: P63085 Mouse
    • SwissProt: P63086 Rat
    • Unigene: 431850 Human
    • Unigene: 196581 Mouse
    • Unigene: 34914 Rat
    see all
  • Alternative names

    • ERK 2 antibody
    • ERK-2 antibody
    • ERT1 antibody
    • Extracellular Signal Regulated Kinase 2 antibody
    • Extracellular signal-regulated kinase 2 antibody
    • MAP kinase 1 antibody
    • MAP kinase 2 antibody
    • MAP kinase isoform p42 antibody
    • MAPK 1 antibody
    • MAPK 2 antibody
    • Mapk1 antibody
    • MAPK2 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
    • P42MAPK antibody
    • PRKM1 antibody
    • PRKM2 antibody
    • protein kinase, mitogen-activated, 1 antibody
    • protein kinase, mitogen-activated, 2 antibody
    • protein tyrosine kinase ERK2 antibody
    see all

Images

  • Western blot - Anti-ERK2 antibody [E460] (ab32081)
    Western blot - Anti-ERK2 antibody [E460] (ab32081)
    All lanes : Anti-ERK2 antibody [E460] (ab32081) at 1/1000 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: 41 kDa
    Observed band size: 41 kDa



    Lanes 1-2: Merged signal (red and green). Green - ab32081 observed at 41 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab32081 Anti-ERK2 antibody [E460] 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. ab32081 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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.

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK2 antibody [E460] (ab32081)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK2 antibody [E460] (ab32081)

    IHC image of ab32081 staining ERK2 in rat pancreas 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 ab32081, 5μg/ml, 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.

  • Western blot - Anti-ERK2 antibody [E460] (ab32081)
    Western blot - Anti-ERK2 antibody [E460] (ab32081)
    All lanes :

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : EKR2 knockout HAP1 cell lysate
    Lane 3 : NIH3T3 cell lysate
    Lane 4 : PC12 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 41 kDa



    ab32081 was shown to specifically react with ERK2 (MAPK1) in wild type HAP1 cells. No band was observed when ERK2 (MAPK1) knockout samples were used. Wild-type and ERK2 (MAPK1) knockout samples were subjected to SDS-PAGE. ab32081 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK2 antibody [E460] (ab32081)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK2 antibody [E460] (ab32081)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ERK2 with ab32081 at 1/400. Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
    Control: PBS only.
    Nuclear counter stain: DAPI.

  • Western blot - Anti-ERK2 antibody [E460] (ab32081)
    Western blot - Anti-ERK2 antibody [E460] (ab32081)
    All lanes : Anti-ERK2 antibody [E460] (ab32081)

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : ERK2 knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 41 kDa



    Lanes 1 - 2: Merged signal (red and green). Green - ab32081 observed at 44 kDa. Red signal from loading control - ab8226 observed at 42 kDa or ab8245, observed at 37 kDa. 

    This western blot image is a comparison between ab32081 and a competitor's top cited rabbit polyclonal antibody.

  • Flow Cytometry (Intracellular) - Anti-ERK2 antibody [E460] (ab32081)
    Flow Cytometry (Intracellular) - Anti-ERK2 antibody [E460] (ab32081)

    Overlay histogram showing HeLa cells stained with ab32081 (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 (ab32081, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluorr® 488 (IgG; H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/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. 

     

     

  • Western blot - Anti-ERK2 antibody [E460] (ab32081)
    Western blot - Anti-ERK2 antibody [E460] (ab32081)
    All lanes : Anti-ERK2 antibody [E460] (ab32081) 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 : 46C (Mouse neural progenitor, selected for Sox1 expression cell line) Whole Cell Lysate
    Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 5 : Recombinant Human ERK1 protein (ab43623) (ab43623)
    Lane 6 : Recombinant Human ERK2 protein (ab43625)

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 41 kDa
    Observed band size: 41 kDa



    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 ab32081 overnight at 4°C. Antibody binding was detected using ab175781 (goat anti-rabbit Alexa Fluor 790) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK2 antibody [E460] (ab32081)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK2 antibody [E460] (ab32081)Image from Emmanuel C et al., PLoS One. 2011 Mar 15;6(3):e17617. doi: 10.1371/journal.pone.0017617.; Fig 2.; March 15, 2011, PLoS ONE 6(3): e17617.

    Immunohistochemical analysis of Human fallopian tissue epithelium, staining ERK2 with ab32081 at 1/50 dilution. Samples were incubated with primary antibody for 1 hour at room temperature. Staining was detected using DAB.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-ERK2 antibody [E460] (ab32081)
    Anti-ERK2 antibody [E460] (ab32081)

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

  • SDS download

  • Datasheet download

    Download

References (31)

Publishing research using ab32081? Please let us know so that we can cite the reference in this datasheet.

ab32081 has been referenced in 31 publications.

  • Li X  et al. Circular RNA UBAP2 promotes the proliferation of prostate cancer cells via the miR-1244/MAP3K2 axis. Oncol Lett 21:486 (2021). PubMed: 33968202
  • Han J  et al. Quantitative proteomic analysis identified differentially expressed proteins with tail/rump fat deposition in Chinese thin- and fat-tailed lambs. PLoS One 16:e0246279 (2021). PubMed: 33529214
  • Shi M  et al. miR-362-3p Targets Orosomucoid 1 to Promote Cell Proliferation, Restrain Cell Apoptosis and Thereby Mitigate Hypoxia/Reoxygenation-Induced Cardiomyocytes Injury. Cardiovasc Toxicol 21:387-398 (2021). PubMed: 33459949
  • Ngo B  et al. Limited Environmental Serine and Glycine Confer Brain Metastasis Sensitivity to PHGDH Inhibition. Cancer Discov 10:1352-1373 (2020). PubMed: 32571778
  • Guo G  et al. Testosterone modulates structural synaptic plasticity of primary cultured hippocampal neurons through ERK - CREB signalling pathways. Mol Cell Endocrinol 503:110671 (2020). PubMed: 31805308
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Flow Cytometry abreview for Anti-ERK2 antibody [E460]

Excellent
Abreviews
Abreviews
abreview image
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
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.

Abcam user community

Verified customer

Submitted Aug 04 2014

Western blot abreview for Anti-ERK2 antibody [E460]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
50000 cells
Gel Running Conditions
Reduced Denaturing (10%)
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
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.

Abcam user community

Verified customer

Submitted Aug 04 2014

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