Overview

  • Product name
    Anti-c-Jun antibody - ChIP Grade
    See all c-Jun primary antibodies
  • Description
    Rabbit polyclonal to c-Jun - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    Antibody detects endogenous levels of c-Jun protein around Serine 243.
  • Tested applications
    Suitable for: ChIP, ICC/IF, IHC-P, IP, WB, ELISA, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey
    Predicted to work with: Chicken, Cow, Pig
  • Immunogen

    Synthetic peptide within Human c-Jun aa 210-259. The exact sequence is proprietary.
    Sequence:

    HLPQQMPVQHPRLQALKEEPQTVPEMPGETPPLSPIDMESQERIKAERKR


    Database link: P05412

  • Positive control
    • ChIP: Human endothelial cells (EA.hy926). IHC-P: Human breast carcinoma tissue. ICC/IF: HepG2 cells. WB: Extracts of HeLa cells. Recombinant Human c-Jun protein. Flow Cytometry: Jurkat cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab31419 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
ChIP Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/50 - 1/100.
IP Use at an assay dependent concentration.
WB 1/500 - 1/1000. Predicted molecular weight: 36 kDa.
ELISA 1/20000.
Flow Cyt 1/300.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Transcription factor that recognizes and binds to the enhancer heptamer motif 5'-TGA[CG]TCA-3'. Promotes activity of NR5A1 when phosphorylated by HIPK3 leading to increased steroidogenic gene expression upon cAMP signaling pathway stimulation. Involved in activated KRAS-mediated transcriptional activation of USP28 in colorectal cancer (CRC) cells (PubMed:24623306). Binds to the USP28 promoter in colorectal cancer (CRC) cells (PubMed:24623306).
  • Sequence similarities
    Belongs to the bZIP family. Jun subfamily.
    Contains 1 bZIP (basic-leucine zipper) domain.
  • Post-translational
    modifications
    Ubiquitinated by the SCF(FBXW7), leading to its degradation. Ubiquitination takes place following phosphorylation, that promotes interaction with FBXW7.
    Phosphorylated by CaMK4 and PRKDC; phosphorylation enhances the transcriptional activity. Phosphorylated by HIPK3. Phosphorylated by DYRK2 at Ser-243; this primes the protein for subsequent phosphorylation by GSK3B at Thr-239. Phosphorylated at Thr-239, Ser-243 and Ser-249 by GSK3B; phosphorylation reduces its ability to bind DNA. Phosphorylated by PAK2 at Thr-2, Thr-8, Thr-89, Thr-93 and Thr-286 thereby promoting JUN-mediated cell proliferation and transformation. Phosphorylated by PLK3 following hypoxia or UV irradiation, leading to increase DNA-binding activity.
    Acetylated at Lys-271 by EP300.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Activator protein 1 antibody
    • AP 1 antibody
    • AP1 antibody
    • cJun antibody
    • Enhancer Binding Protein AP1 antibody
    • Jun Activation Domain Binding Protein antibody
    • JUN antibody
    • Jun oncogene antibody
    • JUN protein antibody
    • Jun proto oncogene antibody
    • JUN_HUMAN antibody
    • JUNC antibody
    • Oncogene JUN antibody
    • p39 antibody
    • Proto oncogene c jun antibody
    • Proto oncogene cJun antibody
    • Proto-oncogene c-jun antibody
    • Transcription Factor AP 1 antibody
    • Transcription factor AP-1 antibody
    • Transcription Factor AP1 antibody
    • V jun avian sarcoma virus 17 oncogene homolog antibody
    • V jun sarcoma virus 17 oncogene homolog (avian) antibody
    • V jun sarcoma virus 17 oncogene homolog antibody
    • V-jun avian sarcoma virus 17 oncogene homolog antibody
    • vJun Avian Sarcoma Virus 17 Oncogene Homolog antibody
    see all

Images

  • ChIP analysis using ab31419 binding c-Jun in human endothelial cells (EA.hy926). Cells were cross-linked for 10 minutes with formaldehyde then incubated with undiluted primary antibody for 10 hours at 4°C in 1x ChIP buffer. Protein binding was detected using real-time PCR.
    Positive control: Position 89340150-89340297 in chromosome 11 (has a validated c-Jun site).
    Negative Control: Igr5 intron 3 (contains no c-Jun binding site).

    See Abreview

  • Paraffin-embedded human breast carcinoma tissue stained for c-Jun with ab31419 at a 1/50 dilution in immunohistochemical analysis.

    Left panel: Untreated.

    Right panel: Pre-incubated with synthesized peptide.

  • ICC/IF image of  HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling c-Jun (green) with ab31419 at 1 µg/ml. The cells were fixed in 4% PFA (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with ab31419 at 1 µg/ml overnight at +4 °C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) ab150077 used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

  • All lanes : Anti-c-Jun antibody - ChIP Grade (ab31419) at 1/500 dilution

    Lane 1 : Extracts of Hela (Human epithelial cell line from cervix adenocarcinoma) cells
    Lane 2 : Extracts of Hela (Human epithelial cell line from cervix adenocarcinoma) cells with immunizing peptide

    Predicted band size: 36 kDa
    Observed band size: 43 kDa (why is the actual band size different from the predicted?)

  • c-Jun was immunoprecipitated from HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate using ab31419 at a 1/500 dilution.

    Lane 1: Control IgG IP in HEK-293T whole cell lysate.

    Lane 2: ab31419 in HEK-293T whole cell lysate.

    Lane 3: c-Jun in HEK-293T whole cell lysate 500 µg (Input).

    For western blotting an HRP-conjugated swine anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab31419 (blue line) at a 1/300 dilution. The cells were prepared as follows: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight. Buffer was 0.1% sodium azide with FBS in phosphate buffered solution. Cells were fixed using paraformaldehyde and permeabilized using Triton X-100 and NP40. Cells were gated by isolating cell population from plot of SSC-A / FSA-A. The secondary antibody used was a FITC-conjugated Goat anti-Rabbit polyclonal, diluted 1/100.

    See Abreview

  • Anti-c-Jun antibody - ChIP Grade (ab31419) at 1/500 dilution + Recombinant Human c-Jun protein (ab54318) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa


    Exposure time: 30 seconds

References

This product has been referenced in:
  • Vincent K  et al. Intracellular mGluR5 plays a critical role in neuropathic pain. Nat Commun 7:10604 (2016). IHC . Read more (PubMed: 26837579) »
  • Hanson RL  et al. Identification of FRA-1 as a novel player in pancreatic cancer in cooperation with a MUC1: ERK signaling axis. Oncotarget 7:39996-40011 (2016). Read more (PubMed: 27220889) »

See all 27 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Na-citrate pH6
Sample
Mouse Tissue sections (Liver, P5)
Specification
Liver, P5
Permeabilization
Yes - Triton 0,05%
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 07 2014

Application
Immunoprecipitation
Immuno-precipitation step
Other - EMSA
Sample
Human Cell lysate - nuclear (renal cancer cells)
Specification
renal cancer cells
Total protein in input
3 µg
Username

Abcam user community

Verified customer

Submitted Mar 14 2014

ChIP

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Detection step
Real-time PCR
Sample
Human Cell lysate - whole cell (EA.hy926 endothelial cells)
Specification
EA.hy926 endothelial cells
Negative control
Igr5 intron 3 contains no c-Jun binding site (Aguilera C et al. Nature. 2011 469: 231-235)
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde (final concentrat
Positive control
Position 89340150-89340297 in chromosome 11 has a validated c-Jun site (ENCODE Project Consortium. PLoS Biol. 2011 9:e1001046
Username

Abcam user community

Verified customer

Submitted Jun 05 2013

Western blot

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Human Cell lysate - whole cell (EA.hy926 endothelial cells)
Specification
EA.hy926 endothelial cells
Treatment
c-Jun siRNA for 48h
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jun 05 2013

Thank you for your enquiry regarding ab31419 and ab47476; and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with these two antibodies.
I would like to reassure ...

Read More

Thank you for contacting us.

I do wish to aplogize for the delays in getting information about this product to you. I have been informded by the originator of this product that the images are using ductal breast carcinoma however as of thi...

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Thank you very much for contacting us.

I just wanted to let you know that I am still working on your inquiry. I have contacted the originator of this product for the information you requested, and I am awaiting a reply.

I can pr...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116)
Loading amount
30 µg
Specification
HCT116
Gel Running Conditions
Non-reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 07 2012

Thank you for your reply.

Unfortunately we do not have another C-Jun antibody that is tested in ChIP. Please let me know how you would like to proceed. We do not currently stock a C-Fos antibody that will work in ChIP but if you would like t...

Read More

Thank you for your reply and with clarifying information.

We have not received any other complaints about this antibody, so I am sorry to hear you have been experiencing problems. It is troubling that it is not working in both ChIP and WB. I...

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1-10 of 21 Abreviews or Q&A

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