Recombinant
RabMAb

Recombinant Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (ab227533)

Rabbit recombinant monoclonal c-Jun (phospho S63) antibody [Y172]. Validated in WB, ELISA, IHC, Dot, ICC/IF and tested in Mouse, Rat, Human.

Overview

  • Product name

    Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free
    See all c-Jun primary antibodies
  • Description

    Rabbit monoclonal [Y172] to c-Jun (phospho S63) - BSA and Azide free
  • 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, Dot blot, ICC/IF, ELISAmore details
    Unsuitable for: Flow Cyt or IP
  • 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

    Ab227533 is the carrier-free version of ab32385. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab227533 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab227533 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 Use at an assay dependent concentration. Detects a band of approximately 42 kDa (predicted molecular weight: 36 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

Dot blot Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • 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
      • 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

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

      Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates, 15 ug
      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.

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

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

       

    • All lanes : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1ug/mL anisomycin for 15 minutes whole cell lysates
      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.

      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



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

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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: c-Jun (pS63) phospho peptide.
      Lane 2: c-Jun non-phospho peptide.
      Exposure time 3 minutes.

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

    • This IHC data was generated using the same anti-c Jun antibody clone, Y172, in a different buffer formulation (cat# ab32385).

      Ab32385, at a 1/50 dilution, staining c-Jun in paraffin embedded human breast carcinoma tissue by Immunohistochemistry.

    References

    This product has been referenced in:

    • 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). Read more (PubMed: 30472438) »
    • 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) »
    See all 37 Publications for this product

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