RecombinantRabMAb

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

Reviews (1)Q&A (1)References (50)
Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • ELISA - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Dot Blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
  • Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y172] to c-Jun (phospho S63)
  • Suitable for: WB, IHC-P, ICC/IF, Dot blot, ELISA
  • Reacts with: Mouse, Human

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

    Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) 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, ICC/IF, Dot blot, ELISAmore 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 whole cell lysate (ab150035). IHC-P: Human breast carcinoma tissue. ICC/IF: A431 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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/IF 1/100 - 1/200.
Dot blot 1/1000.
ELISA 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 (phospho S63) antibody [Y172] (ab32385)
      Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      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
    • Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

      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).

    • Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).

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

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

      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.

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

      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: Human c-Jun (pS63) phospho peptide.
      Lane 2: Human c-Jun non-phospho peptide.
      Exposure time 3 minutes.

    • Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      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?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

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

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

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (50)

    Publishing research using ab32385? Please let us know so that we can cite the reference in this datasheet.

    ab32385 has been referenced in 50 publications.

    • 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
    • Yadav P  et al. Glutathione S-transferasesP1 AA (105Ile) allele increases oral cancer risk, interacts strongly with c-Jun Kinase and weakly detoxifies areca-nut metabolites. Sci Rep 10:6032 (2020). PubMed: 32265484
    • Goodwani S  et al. Dual Leucine Zipper Kinase Is Constitutively Active in the Adult Mouse Brain and Has Both Stress-Induced and Homeostatic Functions. Int J Mol Sci 21:N/A (2020). WB, IHC-FrFl ; Mouse . PubMed: 32659913
    • Ma Z  et al. Inhibitory effect of simvastatin in nasopharyngeal carcinoma cells. Exp Ther Med 17:4477-4484 (2019). PubMed: 31105786
    • Lakkappa N  et al. Soluble epoxide hydrolase inhibitor, APAU, protects dopaminergic neurons against rotenone induced neurotoxicity: Implications for Parkinson's disease. Neurotoxicology 70:135-145 (2019). PubMed: 30472438
    View all Publications for this product

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    Western blot abreview for Anti-c-Jun (phospho S63) antibody [Y172]

     Inconclusive
    Abreviews
    abreview image
    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
    Read More

    Mrs. Katie Elliott

    Verified customer

    Submitted Apr 25 2017

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