Product nameAnti-c-Jun antibody [E254]
See all c-Jun primary antibodies
DescriptionRabbit monoclonal [E254] to c-Jun
Tested applicationsSuitable for: WB, IHC-P, IP, ICC/IFmore details
Unsuitable for: Flow Cyt
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Pig
Synthetic peptide within Human c-Jun aa 1-100 (N terminal). The exact sequence is proprietary.
Epitopeab32137 reacts with an epitope located in the N terminal region of c-Jun.
- WB: NIH 3T3, HeLa cell lysate ICC/IF HeLa IHC-P: Skin carcinoma IP: HEK-293, NIH3T3 cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Dissociation constant (KD)KD = 2.23 x 10 -11 M Learn more about KD
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
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.
|WB||1/1000 - 1/10000. Predicted molecular weight: 36 kDa.|
|IP||Use a concentration of 5 µg/ml.|
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
- AP1 antibody
Anti-c-Jun antibody [E254] (ab32137) at 1/50000 dilution + PC-12(Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
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?
ab32137 staining c-Jun in HeLa cells treated with curcumin (diferuloylmethane) (ab120618), by ICC/IF. Decrease in c-Jun expression correlates with increased concentration of curcumin (diferuloylmethane) as described in literature.
The cells were incubated at 37°C for 4h in media containing different concentrations of ab120618 (curcumin (diferuloylmethane)) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32137 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
All lanes : Anti-c-Jun antibody [E254] (ab32137) at 1/2000 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.
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).
Anti-c-Jun antibody [E254] (ab32137) at 1/2000 dilution + NIH 3T3 cell lysate
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?
Immunohistochemical analysis of c-Jun expression in paraffin embedded skin carcinoma tissue sample, using 1/250 ab32137.
ab32137 (purified) at 1/500 dilution (2.29 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, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST .
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).
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
ICC/IF image of ab32137 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32137, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KD
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 Y et al. Overexpression of Opa interacting protein 5 increases the progression of liver cancer via BMPR2/JUN/CHEK1/RAC1 dysregulation. Oncol Rep 41:2075-2088 (2019). Read more (PubMed: 30816485) »