Recombinant
RabMAb

Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Overview

  • Product name
    Anti-c-Jun (phospho S63) antibody [Y172]
    See all c-Jun primary antibodies
  • Description
    Rabbit monoclonal [Y172] to c-Jun (phospho S63)
  • Host species
    Rabbit
  • Specificity
    The antibody only detects c-Jun phosphorylated on Serine 63 when tested in WB and ICC using specific phospho-treatments. However, in DotBlot and ELISA assays we detected some cross-reactivity with the non-phospho peptide as well. Please refer to the images on the datasheet. The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
  • Tested applications
    Suitable for: WB, IHC-P, ICCmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow
  • Immunogen

    Synthetic peptide within Human c-Jun aa 50-150 (phospho S63). The exact sequence is proprietary.
    Database link: P05412

  • Positive control
    • WB: UV or Anisomycin treated NIH/3T3 or HeLa cell lysate. IHC-P: Human breast carcinoma tissue. ICC/IF: A431 cells.
  • General notes

    A trial size is available to purchase for this antibody.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32385 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 1/1000 - 1/10000. Detects a band of approximately 42 kDa (predicted molecular weight: 36 kDa).
IHC-P 1/250.

See IHC antigen retrieval protocols.

For unpurified use at 1/50 - 1/100

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

ICC 1/100 - 1/200.
  • 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
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling c-Jun with Purified ab32385 at 1:250 dilution (0.46 µg/ml). Heat mediated antigen retrieval was performed using 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.Hematoxylinwas used as a counterstain
    • All lanes : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 0.1 µg/ml (purified)

      Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
      Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 250 ng/ml anisomycin for 30 minutes whole cell lysates
      Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 250 ng/ml anisomycin for 30 minutes whole cell lysates. Then the membrane was incubated with phosphatase

      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



      Blocking and diluting buffer: 5% NFDM/TBST.

    • All lanes : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 0.1 µg/ml (purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1 ug/ml anisomycin for 15 minutes whole cell lysates
      Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1 ug/ml anisomycin for 15 minutes whole cell lysates 15ug. Then the membrane was incubated with phosphatase.

      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



      Blocking and diluting buffer: 5% NFDM/TBST.

    • Lane 1 : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 1/1000 dilution (Unpurified)
      Lanes 2-3 : Human HRPT2 peptide (ab23385) at 1/1000 dilution (Unpurified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with NFDM/TBST
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1ug/mL anisomycin for 15 minutes whole cell lysates with NFDM/TBST
      Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1ug/ml anisomycin for 15 minutes whole cell lysates. Then the membrane was incubated with phosphatase. with NFDM/TBST

      Lysates/proteins at 15 µg per lane.

      Blocking peptides at 5 % per lane.

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

      Predicted band size: 36 kDa

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling c-Jun (phospho S63) with ab32385 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing the expression was increased after treatment with anisomycin (1 µg/ml for 15 minutes), then decreased after treatment with the Lambda Protein Phosphatase treatment 31℃ for 2 hours. The nuclear counter stain is DAPI (blue).
      Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).

    • Antigen pS63:c-Jun (phospho S63); NP:c-Jun non-phospho. Antigen concentration 0.01~1 ng/ml.

      Primary antibody concentration range 0~1000 ng/ml.

      Secondary antibody is an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) used at a 1:2500 dilution.

    • Unpurified ab32385 used at a 1:1000 dilution.
      Secondary antibody is Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) used at a 1:100,000 dilution. Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
      Lane 1: c-Jun (pS63) phospho peptide.
      Lane 2: c-Jun non-phospho peptide.
      Exposure time 3 minutes.

    • All lanes : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 1/10000 dilution (Unpurified)

      Lanes 1 & 3 : Untreated NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate
      Lane 2 : NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate treated with ultraviolet light
      Lane 4 : NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate treated with 25 µg/ml Anisomycin for 15 minutes at 37°C

      Predicted band size: 36 kDa
      Observed band size: 42 kDa (why is the actual band size different from the predicted?)

    • Paraffin-embedded human breast carcinoma tissue stained for c-Jun (phospho S63) with unpurified ab32385 at a 1/50 dilution in immunohistochemical analysis.

    References

    This product has been referenced in:
    • Li X  et al. Hepatic loss of Lissencephaly 1 (Lis1) induces fatty liver and accelerates liver tumorigenesis in mice. J Biol Chem 293:5160-5171 (2018). Read more (PubMed: 29475944) »
    • Luan L & Liang Z Tanshinone IIA protects murine chondrogenic ATDC5 cells from lipopolysaccharide-induced inflammatory injury by down-regulating microRNA-203a. Biomed Pharmacother 103:628-636 (2018). WB . Read more (PubMed: 29679904) »

    See all 23 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Chicken Cell lysate - whole cell (Chicken thrombocyte)
    Gel Running Conditions
    Reduced Denaturing (4-20% gradient)
    Loading amount
    10 µg
    Specification
    Chicken thrombocyte
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Mrs. Katie Elliott

    Verified customer

    Submitted Apr 25 2017

    Merci de nous avoir contactés. Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu. Je me permets de vous transférer le numéro de commande de remplacement gratuit de ce produit par une unité de ab32385 : ...

    Read More

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