Revision 6

#9165Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IP, IHC-P, IF-IC, FC-FP, ChIP, ChIP-seq

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

43, 48

Source/Isotype:

Rabbit IgG

UniProt ID:

#P05412

Entrez-Gene Id:

3725

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:150 - 1:600
Immunofluorescence (Immunocytochemistry) 1:200 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #26325.

Specificity / Sensitivity

c-Jun (60A8) Rabbit mAb detects endogenous levels of total c-Jun protein, regardless of phosphorylation state.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a GST-c-Jun protein corresponding to the amino-terminal sequence of human c-Jun.

Background

c-Jun is a member of the Jun family containing c-Jun, JunB, and JunD, and is a component of the transcription factor activator protein-1 (AP-1). AP-1 is composed of dimers of Fos, Jun, and ATF family members and binds to and activates transcription at TRE/AP-1 elements (reviewed in 1). Extracellular signals, including growth factors, chemokines, and stress, activate AP-1-dependent transcription. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 through SAPK/JNK (reviewed in 2). Knockout studies in mice have shown that c-Jun is essential for embryogenesis (3), and subsequent studies have demonstrated roles for c-Jun in various tissues and developmental processes, including axon regeneration (4), liver regeneration (5), and T cell development (6). AP-1 regulated genes exert diverse biological functions, including cell proliferation, differentiation, and apoptosis, as well as transformation, invasion and metastasis, depending on cell type and context (7-9). Other target genes regulate survival, as well as hypoxia and angiogenesis (8,10). Research studies have implicated c-Jun as a promising therapeutic target for cancer, vascular remodeling, acute inflammation, and rheumatoid arthritis (11,12).

  1. Jochum, W. et al. (2001) Oncogene 20, 2401-12.
  2. Davis, R.J. (2000) Cell 103, 239-52.
  3. Hilberg, F. et al. (1993) Nature 365, 179-81.
  4. Raivich, G. et al. (2004) Neuron 43, 57-67.
  5. Behrens, A. et al. (2002) EMBO J 21, 1782-90.
  6. Riera-Sans, L. and Behrens, A. (2007) J Immunol 178, 5690-700.
  7. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
  8. Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6.
  9. Weiss, C. and Bohmann, D. (2004) Cell Cycle 3, 111-3.
  10. Karamouzis, M.V. et al. (2007) Mol Cancer Res 5, 109-20.
  11. Kim, S. and Iwao, H. (2003) J Pharmacol Sci 91, 177-81.
  12. Dass, C.R. and Choong, P.F. (2008) Pharmazie 63, 411-4.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 6
#9165

c-Jun (60A8) Rabbit mAb

Western Blotting Image 1: c-Jun (60A8) Rabbit mAb Expand Image
使用 c-Jun (60A8) Rabbit mAb #9165对对照型 HeLa 细胞 (lane 1) 或 c-Jun 敲除型 HeLa 细胞 (lane 2) 的提取物进行蛋白印迹分析。c-Jun 敲除型 HeLa 细胞中没有信号,这证实了该抗体对 c-Jun 的特异性。
Western Blotting Image 2: c-Jun (60A8) Rabbit mAb Expand Image
使用 c-Jun (60A8) Rabbit mAb 对未经和已经 UV 处理的 NIH/3T3 和 SK-N-MC 细胞提取物进行进行蛋白质印迹分析。
Western Blotting Image 1: c-Jun (60A8) Rabbit mAb Expand Image

使用  c-Jun (60A8) Rabbit mAb #9165 对经过紫外线(100 mJ/cm2;恢复 2 小时)处理的 HeLa 细胞裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。

No image available
Immunohistochemistry Image 1: c-Jun (60A8) Rabbit mAb Expand Image
使用 c-Jun (60A8) Rabbit mAb 对石蜡包埋的人类星形细胞瘤进行免疫组织化学分析。
Immunohistochemistry Image 2: c-Jun (60A8) Rabbit mAb Expand Image
使用 c-Jun (60A8) Rabbit mAb 对石蜡包埋的人类结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 3: c-Jun (60A8) Rabbit mAb Expand Image
使用 c-Jun (60A8) Rabbit mAb 对石蜡包埋的人类卵巢癌进行免疫组织化学分析。
Immunofluorescence Image 1: c-Jun (60A8) Rabbit mAb Expand Image
使用 c-Jun (60A8) Rabbit mAb(绿色)对 Hela 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 Alexa Fluor®555 phalloidin(红色)进行标记。
Flow Cytometry Image 1: c-Jun (60A8) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 c-Jun (60A8) Rabbit mAb(实线)对 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
Chromatin Immunoprecipitation Image 1: c-Jun (60A8) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对饥饿过夜后以β-NGF #5221 (50ng/ml) 处理 2 小时的 PC-12 细胞中提取的交联染色质,在加入 c-Jun (60A8) Rabbit mAb 后进行染色质免疫沉淀。DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795。图片中显示 Dclk1 内结合,而 Dclk1 是已知的 c-Jun 靶标基因(请参阅含有 ChIP-qPCR 数据的附图)。
Chromatin Immunoprecipitation Image 2: c-Jun (60A8) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对饥饿过夜后以β-NGF #5221 (50ng/ml) 处理 2 小时的 PC-12 细胞中提取的交联染色质,在加入 c-Jun (60A8) Rabbit mAb 后进行染色质免疫沉淀。DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795。结果图显示在染色体2(上图)内的结合,包括 c-Jun 的已知靶标基因 Dclk1(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: c-Jun (60A8) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对饥饿过夜后以 β-NGF #5221 (50ng/ml) 处理 2 小时的 PC-12 细胞中提取的交联染色质,在加入 c-Jun (60A8) Rabbit mAb 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀。使用 SimpleChIP® Rat CCRN4L Promoter Primers #7983、rat DCLK1 promoter primers 和 SimpleChIP® Rat GAPDH Promoter Primers #7964,通过实时 PCR 对富集的 DNA 进行定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。