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RecombinantRabMAb

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

Reviews (4)Q&A (1)References (95)
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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free HRP

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

    Suitable for: WB, IHC-P, IP, ChIPmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Pig

  • 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.

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.

Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 36 kDa.
IHC-P (2)
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 (95)

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

ab32137 has been referenced in 95 publications.

  • Bhattacharya A  et al. MUC1-C Dictates JUN and BAF-Mediated Chromatin Remodeling at Enhancer Signatures in Cancer Stem Cells. Mol Cancer Res 20:556-567 (2022). PubMed: 35022313
  • Brandolini L  et al. Paclitaxel binds and activates C5aR1: A new potential therapeutic target for the prevention of chemotherapy-induced peripheral neuropathy and hypersensitivity reactions. Cell Death Dis 13:500 (2022). PubMed: 35614037
  • Huo W  et al. Triclosan activates c-Jun/miR-218-1-3p/SLC35C1 signaling to regulate cell viability, migration, invasion and inflammatory response of trophoblast cells in vitro. BMC Pregnancy Childbirth 22:470 (2022). PubMed: 35668364
  • Mondal S  et al. PROBER identifies proteins associated with programmable sequence-specific DNA in living cells. Nat Methods 19:959-968 (2022). PubMed: 35927480
  • Zu Y  et al. The roles of CPSF6 in proliferation, apoptosis and tumorigenicity of lung adenocarcinoma. Aging (Albany NY) 14:9300-9316 (2022). PubMed: 36446361
View all Publications for this product

Customer reviews and Q&As

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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
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.

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Verified customer

Submitted Mar 26 2019

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