Recombinant Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E254] to c-Jun - ChIP Grade
- Suitable for: WB, IHC-P, IP, ChIP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-c-Jun antibody [E254] - ChIP Grade
See all c-Jun primary antibodies -
Description
Rabbit monoclonal [E254] to c-Jun - ChIP Grade -
Host species
Rabbit -
Specificity
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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Tested applications
Suitable for: WB, IHC-P, IP, ChIPmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Pig -
Immunogen
Synthetic peptide within Human c-Jun aa 1-100 (N terminal). The exact sequence is proprietary.
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Epitope
ab32137 reacts with an epitope located in the N terminal region of c-Jun. -
Positive control
- WB: NIH 3T3 and HeLa cell lysate. IHC-P: Human lung carcinoma tissue IP: HEK-293 and NIH3T3 cells.
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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
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.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E254 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 36 kDa.
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IHC-P | (2) |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
IP |
1/60.
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ChIP |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 36 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
IP
1/60. |
ChIP
Use at an assay dependent concentration. |
Target
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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
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. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 3725 Human
- Entrez Gene: 16476 Mouse
- Entrez Gene: 396913 Pig
- Entrez Gene: 24516 Rat
- Omim: 165160 Human
- SwissProt: P05412 Human
- SwissProt: P05627 Mouse
- SwissProt: P56432 Pig
see all -
Alternative names
- Activator protein 1 antibody
- AP 1 antibody
- AP-1 antibody
see all
Images
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All lanes : Anti-c-Jun antibody [E254] - ChIP Grade (ab32137) at 1/200 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.
Secondary
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 minutesBlocking 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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling c-Jun with purified ab32137 at 1:100 dilution (1.2 µg/ml). Heat mediated antigen retrieval was performed 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. Hematoxylin was used as a counterstain.
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Anti-c-Jun antibody [E254] - ChIP Grade (ab32137) at 1/5000 dilution + PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Secondary
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 (purified) at 1/60 dilution (20 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, ab32137 at 1/500 dilution (2.29 µg/mL) and
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).
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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
Datasheets and documents
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SDS download
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Datasheet download
References (95)
ab32137 has been referenced in 95 publications.
- Bhattacharya A et al. MUC1-C Dictates JUN and BAF-Mediated Chromatin Remodeling at Enhancer Signatures in Cancer Stem Cells. Mol Cancer Res 20:556-567 (2022). PubMed: 35022313
- Brandolini L et al. Paclitaxel binds and activates C5aR1: A new potential therapeutic target for the prevention of chemotherapy-induced peripheral neuropathy and hypersensitivity reactions. Cell Death Dis 13:500 (2022). PubMed: 35614037
- Huo W et al. Triclosan activates c-Jun/miR-218-1-3p/SLC35C1 signaling to regulate cell viability, migration, invasion and inflammatory response of trophoblast cells in vitro. BMC Pregnancy Childbirth 22:470 (2022). PubMed: 35668364
- Mondal S et al. PROBER identifies proteins associated with programmable sequence-specific DNA in living cells. Nat Methods 19:959-968 (2022). PubMed: 35927480
- Zu Y et al. The roles of CPSF6 in proliferation, apoptosis and tumorigenicity of lung adenocarcinoma. Aging (Albany NY) 14:9300-9316 (2022). PubMed: 36446361