Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP693Y] to c-Jun
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Rat, Human
Product nameAnti-c-Jun antibody [EP693Y]
See all c-Jun primary antibodies
DescriptionRabbit monoclonal [EP693Y] to c-Jun
SpecificityPBS only lot tested.
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Rat, Human
Predicted to work with: Mouse
Synthetic peptide within Human c-Jun aa 300-400 (C terminal). The exact sequence is proprietary.
- WB: PC-12 and NIH/3T3 whole cell lysate. HEK-293 cell lysate. IHC-P: Human colon carcinoma tissue. ICC/IF: HeLa cells. ICC/IF KO: HEK293 cells (HEK293-JUN KO cells used as a negative cell line).
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab40766 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/5000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/100 - 1/250.
Signal can be observed in cells fixed with either methanol or paraformaldehyde.
FunctionTranscription 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 similaritiesBelongs to the bZIP family. Jun subfamily.
Contains 1 bZIP (basic-leucine zipper) domain.
modificationsUbiquitinated 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.
- Information by UniProt
- Activator protein 1 antibody
- AP 1 antibody
- AP-1 antibody
ab40766 staining c-Jun in wild-type HEK293 cells (top panel) and c-Jun knockout HEK293 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab40766 at 1/250 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).
All lanes : Anti-c-Jun antibody [EP693Y] (ab40766) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : Jun knockout HEK-293 whole cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 39 kDa
Lanes 1 - 2: Merged signal (red and green). Green - ab40766 observed at 35 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab40766 was shown to specifically react with Jun in wild-type HEK-293 cells as signal was lost in Jun knockout cells. Wild-type and Jun knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab40766 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-c-Jun antibody [EP693Y] (ab40766) at 0.212 µg/ml
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 39 kDa
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: Lane 1- 41 seconds
Lane 2 - 1 second
Anti-c-Jun antibody [EP693Y] (ab40766) at 1/5000 dilution + 293 cell lysate at 10 µg
Predicted band size: 39 kDa
Observed band size: 39 kDa
Tissue: Paraffin-embedded human colon carcinoma.
Staining: ab40766 at 1/100 dilution
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunofluorescent staining of HeLa cells
ab40766 at 1/100 dilution
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.
ab40766 has been referenced in 8 publications.
- Kong YL et al. Silencing of the MEKK2/MEKK3 Pathway Protects against Spinal Cord Injury via the Hedgehog Pathway and the JNK Pathway. Mol Ther Nucleic Acids 17:578-589 (2019). PubMed: 31382189
- Cao Y et al. CENPO expression regulates gastric cancer cell proliferation and is associated with poor patient prognosis. Mol Med Rep 20:3661-3670 (2019). PubMed: 31485675
- Xu E et al. Hypoxia-Inducible Factor 1A Upregulates HMGN5 by Increasing the Expression of GATA1 and Plays a Role in Osteosarcoma Metastasis. Biomed Res Int 2019:5630124 (2019). PubMed: 31930127
- Fernandes KA et al. Role of SARM1 and DR6 in retinal ganglion cell axonal and somal degeneration following axonal injury. Exp Eye Res 171:54-61 (2018). PubMed: 29526794
- Syc-Mazurek SB et al. JUN is important for ocular hypertension-induced retinal ganglion cell degeneration. Cell Death Dis 8:e2945 (2017). IHC ; Mouse . PubMed: 28726785
- Fernandes KA et al. Novel axon projection after stress and degeneration in the Dscam mutant retina. Mol Cell Neurosci 71:1-12 (2016). PubMed: 26691152
- Mac Nair CE et al. Tumor necrosis factor alpha has an early protective effect on retinal ganglion cells after optic nerve crush. J Neuroinflammation 11:194 (2014). PubMed: 25407441
- Chen YJ et al. Global assessment of Antrodia cinnamomea-induced microRNA alterations in hepatocarcinoma cells. PLoS One 8:e82751 (2013). WB ; Human . PubMed: 24358224