Anti-c-Jun抗体- ChIP Grade (ab31419)

概述

  • 产品名称
    Anti-c-Jun抗体- ChIP Grade
    参阅全部 c-Jun 一抗
  • 描述
    兔多克隆抗体to c-Jun - ChIP Grade
  • 特异性
    Antibody detects endogenous levels of c-Jun protein around Serine 243.
  • 经测试应用
    适用于: ChIP, ICC/IF, IHC-P, IP, WB, ELISA, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, African green monkey
    预测可用于: Chicken, Cow, Pig
  • 免疫原

    Synthetic peptide corresponding to Human c-Jun aa 210-259. Range of aa 210 - 259
    Sequence:

    H LPQQMPVQHP RLQALKEEPQ TVPEMPGETP PLSPIDMESQ ERIKAERKR


    Database link: P05412

  • 阳性对照
    • Extracts of Hela cells and breast carcinoma tissue. HepG2 cells. 3T3 or Raw cells.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能
    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).
  • 序列相似性
    Belongs to the bZIP family. Jun subfamily.
    Contains 1 bZIP (basic-leucine zipper) domain.
  • 翻译后修饰
    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.
  • 细胞定位
    Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • ChIP analysis using ab31419 binding c-Jun in Human endothelial cells (EA.hy926). Cells were cross-linked for 10 minutes with formaldehyde. Samples were 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

  • Immunohistochemical analysis of paraffin embedded Human breast carcinoma labeling c-Jun with ab31419 at 1/50.

    Right image: Antibody pre-incubated with synthesized peptide.


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa


    Exposure time : 30 seconds
  • ICC/IF image of  HepG2 cells labeling c-Jun (green) with ab31419 at 1 µg/ml. The cells were 4% PFA 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 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 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 1.43 µM.

  • All lanes : Anti-c-Jun antibody - ChIP Grade (ab31419)

    Lane 1 : Extracts of Hela cells.
    Lane 2 : Extracts of Hela cells. Antibody pre-incubated with synthesized peptide.


    Predicted band size : 36 kDa
    Observed band size : 43 kDa (why is the actual band size different from the predicted?)
  • Overlay histogram showing Jurkat cells stained with ab31419 (blue line). 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. ab31419 was diluted 1/300 and incubated at 4°C for 24 hours overnight. The secondary antibody used was a FITC-conjugated Goat anti-Rabbit polyclonal, diluted 1/100.

    See Abreview

文献

This product has been referenced in:
  • Vincent K  et al. Intracellular mGluR5 plays a critical role in neuropathic pain. Nat Commun 7:10604 (2016). IHC . Read more (PubMed: 26837579) »
  • Hanson RL  et al. Identification of FRA-1 as a novel player in pancreatic cancer in cooperation with a MUC1: ERK signaling axis. Oncotarget 7:39996-40011 (2016). Read more (PubMed: 27220889) »

See all 27 Publications for this product

客户评价及客户问答

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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 Jun 05 2013

Thank you for your enquiry regarding ab31419 and ab47476; and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with these two antibodies.
I would like to reassure ...

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Thank you for contacting us.

I do wish to aplogize for the delays in getting information about this product to you. I have been informded by the originator of this product that the images are using ductal breast carcinoma however as of thi...

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Thank you very much for contacting us.

I just wanted to let you know that I am still working on your inquiry. I have contacted the originator of this product for the information you requested, and I am awaiting a reply.

I can pr...

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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
Username

Abcam user community

Verified customer

提交于 Nov 07 2012

Thank you for your reply.

Unfortunately we do not have another C-Jun antibody that is tested in ChIP. Please let me know how you would like to proceed. We do not currently stock a C-Fos antibody that will work in ChIP but if you would like t...

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Thank you for your reply and with clarifying information.

We have not received any other complaints about this antibody, so I am sorry to hear you have been experiencing problems. It is troubling that it is not working in both ChIP and WB. I...

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1-10 of 21 Abreviews or Q&A

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

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