製品の概要

  • 製品名
    Anti-c-Jun antibody - ChIP Grade
    c-Jun 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to c-Jun - ChIP Grade
  • 由来種
    Rabbit
  • 特異性
    Antibody detects endogenous levels of c-Jun protein around Serine 243.
  • アプリケーション
    適用あり: ChIP, ICC/IF, IHC-P, IP, WB, ELISA, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Human, African green monkey
    交差が予測される動物種: Chicken, Cow, Pig
  • 免疫原

    Synthetic peptide within Human c-Jun aa 210-259. The exact sequence is proprietary.
    Sequence:

    HLPQQMPVQHPRLQALKEEPQTVPEMPGETPPLSPIDMESQERIKAERKR


    Database link: P05412

  • ポジティブ・コントロール
    • ChIP: Human endothelial cells (EA.hy926). IHC-P: Human breast carcinoma tissue. ICC/IF: HepG2 cells. WB: Extracts of HeLa cells. Recombinant Human c-Jun protein. Flow Cytometry: Jurkat cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab31419 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ChIP Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/50 - 1/100.
IP Use at an assay dependent concentration.
WB 1/500 - 1/1000. Predicted molecular weight: 36 kDa.
ELISA 1/20000.
Flow Cyt 1/300.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能
    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).
  • 配列類似性
    Belongs to the bZIP family. Jun subfamily.
    Contains 1 bZIP (basic-leucine zipper) domain.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Activator protein 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

画像

  • ChIP analysis using ab31419 binding c-Jun in human endothelial cells (EA.hy926). Cells were cross-linked for 10 minutes with formaldehyde then incubated with undiluted primary antibody for 10 hours at 4°C in 1x ChIP buffer. Protein binding was detected using real-time PCR.
    Positive control: Position 89340150-89340297 in chromosome 11 (has a validated c-Jun site).
    Negative Control: Igr5 intron 3 (contains no c-Jun binding site).

    See Abreview

  • Paraffin-embedded human breast carcinoma tissue stained for c-Jun with ab31419 at a 1/50 dilution in immunohistochemical analysis.

    Left panel: Untreated.

    Right panel: Pre-incubated with synthesized peptide.

  • ICC/IF image of  HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling c-Jun (green) with ab31419 at 1 µg/ml. The cells were fixed in 4% PFA (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with ab31419 at 1 µg/ml overnight at +4 °C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) ab150077 used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

  • All lanes : Anti-c-Jun antibody - ChIP Grade (ab31419) at 1/500 dilution

    Lane 1 : Extracts of Hela (Human epithelial cell line from cervix adenocarcinoma) cells
    Lane 2 : Extracts of Hela (Human epithelial cell line from cervix adenocarcinoma) cells with immunizing peptide

    Predicted band size: 36 kDa
    Observed band size: 43 kDa (why is the actual band size different from the predicted?)

  • c-Jun was immunoprecipitated from HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate using ab31419 at a 1/500 dilution.

    Lane 1: Control IgG IP in HEK-293T whole cell lysate.

    Lane 2: ab31419 in HEK-293T whole cell lysate.

    Lane 3: c-Jun in HEK-293T whole cell lysate 500 µg (Input).

    For western blotting an HRP-conjugated swine anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab31419 (blue line) at a 1/300 dilution. The cells were prepared as follows: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight. Buffer was 0.1% sodium azide with FBS in phosphate buffered solution. Cells were fixed using paraformaldehyde and permeabilized using Triton X-100 and NP40. Cells were gated by isolating cell population from plot of SSC-A / FSA-A. The secondary antibody used was a FITC-conjugated Goat anti-Rabbit polyclonal, diluted 1/100.

    See Abreview

  • Anti-c-Jun antibody - ChIP Grade (ab31419) at 1/500 dilution + Recombinant Human c-Jun protein (ab54318) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa


    Exposure time: 30 seconds

参考文献

This product has been referenced in:
  • Britton E  et al. Open chromatin profiling identifies AP1 as a transcriptional regulator in oesophageal adenocarcinoma. PLoS Genet 13:e1006879 (2017). Read more (PubMed: 28859074) »
  • El-Makakey AM  et al. Comparative study of the efficacy of pulsed electromagnetic field and low level laser therapy on mitogen-activated protein kinases. Biochem Biophys Rep 9:316-321 (2017). Read more (PubMed: 28956019) »

See all 31 Publications for this product

レビューと Q&A

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Na-citrate pH6
Sample
Mouse Tissue sections (Liver, P5)
Specification
Liver, P5
Permeabilization
Yes - Triton 0,05%
Fixative
Paraformaldehyde
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投稿 Oct 07 2014

Application
Immunoprecipitation
Immuno-precipitation step
Other - EMSA
Sample
Human Cell lysate - nuclear (renal cancer cells)
Specification
renal cancer cells
Total protein in input
3 µg
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投稿 Mar 14 2014

ChIP

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Detection step
Real-time PCR
Sample
Human Cell lysate - whole cell (EA.hy926 endothelial cells)
Specification
EA.hy926 endothelial cells
Negative control
Igr5 intron 3 contains no c-Jun binding site (Aguilera C et al. Nature. 2011 469: 231-235)
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde (final concentrat
Positive control
Position 89340150-89340297 in chromosome 11 has a validated c-Jun site (ENCODE Project Consortium. PLoS Biol. 2011 9:e1001046
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投稿 Jun 05 2013

Western blot

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Human Cell lysate - whole cell (EA.hy926 endothelial cells)
Specification
EA.hy926 endothelial cells
Treatment
c-Jun siRNA for 48h
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Jun 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116)
Loading amount
30 µg
Specification
HCT116
Gel Running Conditions
Non-reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Nov 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Jurkat Cells)
Specification
Jurkat Cells
Preparation
Cell harvesting/tissue preparation method: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight.
Sample buffer: Buffer composition at analysis was 0.1% sodium azide with FBS in phosphate buffered solution.
Fixation
Paraformaldehyde
Permeabilization
Yes - Triton X-100 and NP40
Gating Strategy
Isolate cell population from plot of SSC-A / FSA-A
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投稿 Sep 05 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (HeLa)
Loading amount
100 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 Dec 12 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
500 µg
Specification
293T
Immuno-precipitation step
Protein A
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投稿 Oct 30 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Cos-7 cell line)
Loading amount
30 µg
Specification
Cos-7 cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
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投稿 Sep 20 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3 cell line)
Loading amount
30 µg
Specification
3T3 cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
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投稿 Sep 20 2007

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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