Recombinant
RabMAb

Recombinant Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)

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Overview

  • Product name

    Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95]
    See all JNK1+JNK2+JNK3 primary antibodies

  • Description

    Rabbit monoclonal [EPR18841-95] to JNK1+JNK2+JNK3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human JNK1+JNK2+JNK3 aa 150-400. The exact sequence is proprietary. SwissProt ID: P45984 = JNK2 and P53779 = JNK3.
    Database link: P45983

  • Positive control

    • WB: Human JNK1, JNK2 and JNK3 full length recombinant proteins; Human fetal liver, fetal heart and fetal kidney lysates; Jurkat, HeLa, K562, MCF7, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, kidney and spleen lysates. ICC/IF: HeLa and NIH/3T3 cell lines. Flow Cyt: Jurkat and HeLa cell lines. IP: HeLa whole cell lysate.

  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab208035 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 1/2000. Detects a band of approximately 46-54 kDa (predicted molecular weight: 48, 53 kDa).
ICC/IF 1/1000.
Flow Cyt 1/100.
IP 1/40.

Target

  • Function

    Serine/threonine-protein kinase involved in various processes such as cell proliferation, differentiation, migration, transformation and programmed cell death. Extracellular stimuli such as proinflammatory cytokines or physical stress stimulate the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) signaling pathway. In this cascade, two dual specificity kinases MAP2K4/MKK4 and MAP2K7/MKK7 phosphorylate and activate MAPK8/JNK1. In turn, MAPK8/JNK1 phosphorylates a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 and ATF2 and thus regulates AP-1 transcriptional activity. Phosphorylates the replication licensing factor CDT1, inhibiting the interaction between CDT1 and the histone H4 acetylase HBO1 to replication origins. Loss of this interaction abrogates the acetylation required for replication initiation. Promotes stressed cell apoptosis by phosphorylating key regulatory factors including p53/TP53 and Yes-associates protein YAP1. In T-cells, MAPK8 and MAPK9 are required for polarized differentiation of T-helper cells into Th1 cells. Contributes to the survival of erythroid cells by phosphorylating the antagonist of cell death BAD upon EPO stimulation. Mediates starvation-induced BCL2 phosphorylation, BCL2 dissociation from BECN1, and thus activation of autophagy. Phosphorylates STMN2 and hence regulates microtubule dynamics, controlling neurite elongation in cortical neurons. In the developing brain, through its cytoplasmic activity on STMN2, negatively regulates the rate of exit from multipolar stage and of radial migration from the ventricular zone. Phosphorylates several other substrates including heat shock factor protein 4 (HSF4), the deacetylase SIRT1, ELK1, or the E3 ligase ITCH.
    JNK1 isoforms display different binding patterns: beta-1 preferentially binds to c-Jun, whereas alpha-1, alpha-2, and beta-2 have a similar low level of binding to both c-Jun or ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms.

  • 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-183 and Tyr-185 by MAP2K7 and MAP2K4, which activates the enzyme. Phosphorylated by TAOK2.

  • Cellular localization

    Cytoplasm. Nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • C Jun kinase 2 antibody
    • c Jun N terminal kinase 1 antibody
    • c Jun N terminal kinase 2 antibody
    • c Jun N terminal kinase 3 antibody
    • c-Jun N-terminal kinase 1 antibody
    • JNK 46 antibody
    • JNK 55 antibody
    • JNK antibody
    • JNK-46 antibody
    • JNK1 antibody
    • JNK1+2+3 antibody
    • JNK1/2/3 antibody
    • JNK1A2 antibody
    • JNK2 antibody
    • JNK21B1/2 antibody
    • JNK2A antibody
    • JNK2ALPHA antibody
    • JNK2B antibody
    • JNK2BETA antibody
    • JNK3 alpha protein kinase antibody
    • JNK3 antibody
    • JNK3A antibody
    • Jun kinase antibody
    • JUN N terminal kinase antibody
    • MAP kinase 10 antibody
    • MAP kinase 8 antibody
    • MAP kinase 9 antibody
    • MAP kinase p49 3F12 antibody
    • MAPK 10 antibody
    • MAPK 8 antibody
    • MAPK 9 antibody
    • MAPK10 antibody
    • mapk8 antibody
    • MAPK9 antibody
    • Mitogen activated protein kinase 10 antibody
    • Mitogen activated protein kinase 8 antibody
    • Mitogen activated protein kinase 8 isoform JNK1 alpha1 antibody
    • Mitogen activated protein kinase 8 isoform JNK1 beta2 antibody
    • Mitogen activated protein kinase 9 antibody
    • Mitogen-activated protein kinase 8 antibody
    • MK08_HUMAN antibody
    • p493F12 antibody
    • p54a antibody
    • p54aSAPK antibody
    • p54bSAPK antibody
    • PRKM10 antibody
    • PRKM8 antibody
    • PRKM9 antibody
    • SAPK antibody
    • SAPK(beta) antibody
    • SAPK1 antibody
    • SAPK1a antibody
    • SAPK1b antibody
    • SAPK1c antibody
    • Stress activated protein kinase 1 antibody
    • Stress activated protein kinase 1a antibody
    • Stress activated protein kinase 1b antibody
    • Stress activated protein kinase 1c antibody
    • Stress activated protein kinase beta antibody
    • Stress activated protein kinase JNK1 antibody
    • Stress activated protein kinase JNK2 antibody
    • Stress activated protein kinase JNK3 antibody
    • Stress-activated protein kinase 1 antibody
    • Stress-activated protein kinase 1c antibody
    • Stress-activated protein kinase JNK1 antibody
    see all

Images

  • Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution

    Lane 1 : Human JNK1 full length recombinant protein
    Lane 2 : Human JNK2 full length recombinant protein
    Lane 3 : Human JNK3 full length recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 48, 53 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 & 2: 5 seconds; Lane 3: 15 seconds.

    Human JNK1 full length recombinant protein contains aa1-427 with a His-Tag®. Human JNK2 full length recombinant protein contains aa1-424 with a His-Tag®. Human JNK3 full length recombinant protein contains aa1-464 with a His-Tag®.

    All three recombinant human full length proteins were made in-house.

  • Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 48, 53 kDa
    Observed band size: 46-54 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 48, 53 kDa
    Observed band size: 46-54 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 48, 53 kDa
    Observed band size: 46-54 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 4 seconds; Lane 2,3 & 4: 30 seconds; Lane 5: 15 seconds; Lane 6 & 7: 3 minutes.

  • Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Western blot - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 48, 53 kDa
    Observed band size: 46-54 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Immunocytochemistry/ Immunofluorescence - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasm and weak nuclear staining on HeLa cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab208035 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Immunocytochemistry/ Immunofluorescence - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasm and nuclear staining on NIH/3T3 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab208035 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Flow Cytometry - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Flow Cytometry - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)

    Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

    Goat anti-Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

  • Flow Cytometry - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Flow Cytometry - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

    Goat anti-Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
    Immunoprecipitation - Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)

    JNK1+JNK2+JNK3 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab208035 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab208035 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10ug (Input).

    Lane 2: ab208035 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab208035 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

References

This product has been referenced in:

  • Li W  et al. Berberine suppresses IL-33-induced inflammatory responses in mast cells by inactivating NF-?B and p38 signaling. Int Immunopharmacol 66:82-90 (2019). Read more (PubMed: 30445310) »
  • He Z  et al. Inhibition of fowl adenovirus serotype 4 replication in Leghorn male hepatoma cells by SP600125 via blocking JNK MAPK pathway. Vet Microbiol 228:45-52 (2019). Read more (PubMed: 30593379) »
See all 22 Publications for this product

Publishing research using ab208035? Please let us know so that we can cite the reference in this datasheet.

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Western blot abreview for Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat hepatocytes)
Gel Running Conditions
Reduced Denaturing (9%)
Loading amount
20 µg
Specification
Rat hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Read More

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Submitted Jun 03 2016

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