Recombinant
RabMAb

Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956)

Overview

  • Product name
    Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693]
    See all JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) primary antibodies
  • Description
    Rabbit monoclonal [EPR5693] to JNK1 + JNK2 + JNK3 (phospho T183+T183+T221)
  • Specificity
    ab124956 will detect will detect JNK1 (pT183), JNK2 (pT183) and JNK3 (pT221).
  • Tested applications
    Suitable for: Flow Cyt, WB, IP, IHC-P, ICC, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human JNK1 + JNK2 + JNK3 (phospho T183+T183+T221).

  • Positive control
    • NIH 3T3 cell lysates treated with Anisomycin; Human brain tissue.
  • 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 ab124956 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 1/100.

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

WB 1/1000 - 1/10000. Detects a band of approximately 46-54 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes)
ICC 1/50 - 1/100.
ICC/IF Use at an assay dependent concentration.

Target

Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] images

  • All lanes : Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956) at 1/1000 dilution

    Lane 1 : NIH 3T3 cell lysate, untreated
    Lane 2 : NIH 3T3 cell lysate, treated with Anisomycin

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat anti-Rabbit HRP at 1/2000 dilution

    Secondary antibody - goat anti-rabbit HRP (ab6721)

  • Dot blot analysis of JNK1/2/3 (pT183 + pT183 + pT221) peptide (Lane 1) and JNK1/2/3 non-phospho peptide (Lane 2) labelling JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) with ab124956 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunocytochemistry/Immunofluorescence analysis of untreated, Anisomycin treated and Anisomycin + LP treated NIH/3T3 cells labelling JNK1 + JNK2 + JNK3 (phospho T183 + T183 + T221) with ab124956 at a dilution of 1/100 (left) and JNK1 + JNK2 + JNK3 with ab179461 at a dilution of 1/250 (right).

    Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    The image shows increased nuclear staining after Anisomycin (250ng/ml, 30min) treatment on NIH3T3 cells. The LP treatment decreased the increased nuclear staining caused by Anisomycin.

    ab179461 was used as a Pan control for ab124956. The results showed cytoplasmic staining on untreated, Anisomycin and Anisomycin + LP treated NIH3T3 cells.

  • ab124956, at 1/100 dilution staining JNK1+JNK2+JNK3 in paraffin-embedded Human brain tissue, by Immunohistochemistry.
  • Overlay histogram showing HeLa cells stained with ab124956 (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 (ab124956, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor® 488 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.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956)

This product has been referenced in:
  • Wang H  et al. PDK2 promotes chondrogenic differentiation of mesenchymal stem cells by upregulation of Sox6 and activation of JNK/MAPK/ERK pathway. Braz J Med Biol Res 50:e5988 (2017). WB ; Rat . Read more (PubMed: 28225870) »
  • Wang B  et al. IL-34 Upregulated Th17 Production through Increased IL-6 Expression by Rheumatoid Fibroblast-Like Synoviocytes. Mediators Inflamm 2017:1567120 (2017). WB ; Human . Read more (PubMed: 28659662) »

See all 12 Publications for this product

Product Wall

Application
Western blot
Sample
Mouse Tissue lysate - whole (retina)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
40 µg
Specification
retina
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
Username

Abcam user community

Verified customer

Submitted Jun 10 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Mouse Tissue sections (retina)
Specification
retina
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 10 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116 colon cancer cell line)
Loading amount
30 µg
Specification
HCT116 colon cancer cell line
Treatment
DMSO control and 10 µM Irinotecan for 16 hrs
Gel Running Conditions
Reduced Denaturing (13%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Feb 22 2013

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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