RecombinantRabMAb

Recombinant Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)

Reviews (2)Q&A (1)References (64)
Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
  • Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
  • Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
  • ChIP - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
  • Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
  • OI-RD Scanning - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
  • Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E254] to c-Jun - ChIP Grade
  • Suitable for: WB, IHC-P, IP, ChIP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-c-Jun antibody [E254] - ChIP Grade
    See all c-Jun primary antibodies

  • Description

    Rabbit monoclonal [E254] to c-Jun - ChIP Grade

  • Host species

    Rabbit

  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested Applications & Species

    Application Species
    ChIP
    Rat
    IHC-P
    Human
    IP
    Mouse
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human c-Jun aa 1-100 (N terminal). The exact sequence is proprietary.

  • Epitope

    ab32137 reacts with an epitope located in the N terminal region of c-Jun.

  • Positive control

    • WB: NIH 3T3 and HeLa cell lysate. IHC-P: Human lung carcinoma tissue IP: HEK-293 and NIH3T3 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32137 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ChIP
Rat
IHC-P
Human
IP
Mouse
Human
WB
Mouse
Rat
Human
All applications
Pig
Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 36 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
IP
1/60.
ChIP
Use at an assay dependent concentration.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 36 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
IP
1/60.
ChIP
Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt.

    • 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

      see all

    • Alternative names

      • Activator protein 1 antibody
      • AP 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

    • Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      All lanes : Anti-c-Jun antibody [E254] - ChIP Grade (ab32137) at 1/200 dilution

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in 1% SDS Hot lysis method.
      Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates prepared in RIPA lysis method
      Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates prepared in 1% SDS Hot lysis method 15ug.

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 36 kDa
      Observed band size: 39 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking and diluting buffer: 5% NFDM/TBST

      For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling c-Jun with purified ab32137 at 1:100 dilution (1.2 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Anti-c-Jun antibody [E254] - ChIP Grade (ab32137) at 1/5000 dilution + PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

    • Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)

      ab32137 (purified) at 1/60 dilution (20 µg/mL) immunoprecipitating c-Jun in HEK-293 whole cell lysate.
      Lane 2 (+): HEK-293(Human embronic kidney epithelial cell) whole cell lysate 10 µg
      Lane 3 (-): ab32137 & HEK-293 whole cell lysateRabbit monoclonal IgG (ab172730) instead of ab32137 in HEK-293 whole cell lysate
      For western blotting, ab32137 at 1/500 dilution (2.29 µg/mL) and
      VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM /TBST .

    • ChIP - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      ChIP - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Chromatin was prepared from PC-12 (starve overnight) + NGF ( 50 ng/ml 2h) cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab32137 (blue), and 20µl of protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5µg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    • Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      c-Jun was immunoprecipitated using 0.5mg NIH3T3 whole cell extract, 5µg of Rabbit polyclonal to c-Jun and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
      The antibody was incubated under agitation with Protein G beads for 10min, NIH3T3 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32137.
      Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
      Band: 45kDa; c-Jun
    • OI-RD Scanning - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      OI-RD Scanning - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
      Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)

    References (64)

    Publishing research using ab32137? Please let us know so that we can cite the reference in this datasheet.

    ab32137 has been referenced in 64 publications.

    • Liu R  et al. miR-34a targets PAI-1 to regulate urinary microalbumin and renal function in hypertensive mice. Eur J Med Res 25:3 (2020). PubMed: 32178735
    • Li S  et al. Fugan Wan alleviates hepatic fibrosis by inhibiting ACE/Ang II/AT-1R signaling pathway and enhancing ACE2/Ang 1-7/Mas signaling pathway in hepatic fibrosis rat models. Am J Transl Res 12:592-601 (2020). PubMed: 32194907
    • Han M  et al. Abnormality of m6A mRNA Methylation Is Involved in Alzheimer's Disease. Front Neurosci 14:98 (2020). PubMed: 32184705
    • Sun Y  et al. Notoginsenoside R1 alleviates lipopolysaccharide-triggered PC-12 inflammatory damage via elevating microRNA-132. Artif Cells Nanomed Biotechnol 47:1808-1814 (2019). PubMed: 31062615
    • Wan Z  et al. MEIS2 promotes cell migration and invasion in colorectal cancer. Oncol Rep 42:213-223 (2019). PubMed: 31115559
    View all Publications for this product

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-c-Jun antibody [E254]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (IM9 cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin Gel, semi dry transfer on PVDF membrane)
    Loading amount
    25 µg
    Specification
    IM9 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 26 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-c-Jun antibody [E254] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (AML-193)
    Permeabilization
    Yes - 0.1% Tritonx-100 10min
    Specification
    AML-193
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 12 2017

    Question

    about antibodies to be used in ChIP

    Read More
    Answer

    Thank you very much for your call this morning.
    As discussed, I would be happy to provide you with our testing discount to test one of our anti cFos or cJun antibodies in ChIP.
    This morning we discussed the antibodies anti cJun ab32137, anti cFos ab7963 and anti cFos ab129361.
    https://www.abcam.com/c-Jun-antibody-E254-ChIP-Grade-ab32137.html https://www.abcam.com/c-Fos-antibody-ChIP-Grade-ab7963.html
    https://www.abcam.com/c-Fos-antibody-2G2-ab129361.html
    The testing discount I can offer for each of these antiboides is a discount off a future purchase if you buy one of these antibodies now, test it in ChIP and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.
    If you are interested in this offer, please follow these steps:
    1. Reply to this e-mail to let me know which antibodies you would like to test in ChIP. I will then send a discount code for each of the antibodies. This code must be issued before purchasing the antibodies, so please wait for my reply before ordering.
    2. Purchase the antibodies either by phone, fax, or online (www.abcam.com).
    3. Test it in ChIP.
    4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.
    5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.
    We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if an antibody turns out to be unsuitable for ChIP, you will still receive the discount on your next purchase after your Abreview has been submitted.
    Please let me know if you have any questions about this offer and I would be happy to help you further.
    The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com