Revision 11

#9139Store 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-Bond, IHC-P, IF-IC, FC-FP, ChIP, C&R

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

79, 86

Source/Isotype:

Mouse IgG2a kappa

UniProt ID:

#P40763

Entrez-Gene Id:

6774

Product Information

Product Usage Information

For optimal ChIP 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.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:200
IHC Leica Bond 1:300 - 1:1200
Immunohistochemistry (Paraffin) 1:300 - 1:1200
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400
Chromatin IP 1:100
CUT&RUN 1:100

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 #83541.

Specificity / Sensitivity

Stat3 (124H6) Mouse mAb detects endogenous levels of total Stat3 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide centered around amino acid Gln692 of human Stat3.

Background

The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Research studies have shown that Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).

  1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
  2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
  3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
  4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
  5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
  6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
  7. Ihle, J.N. (1995) Nature 377, 591-4.
  8. Wen, Z. et al. (1995) Cell 82, 241-50.
  9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
  10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

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 nonfat dry milk, 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-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP C&R: CUT&RUN

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.
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 11
#9139

Stat3 (124H6) Mouse mAb

Western Blotting Image 1: Stat3 (124H6) Mouse mAb Expand Image
使用 Stat3 (124H6) Mouse mAb #9139(上图)或 β-Actin (8H10D10) Mouse mAb #3700(下图)对各种人细胞系的提取物进行蛋白质印迹法分析。
Western Blotting Image 2: Stat3 (124H6) Mouse mAb Expand Image
使用 Stat3 (124H6) Mouse mAb #9139(上图)或 β-Actin (8H10D10) Mouse mAb #3700(下图)对不同啮齿动物和猴子样本的提取物进行蛋白质印迹法分析。
Western Blotting Image 1: Stat3 (124H6) Mouse mAb Expand Image
使用 Stat3 (124H6) Mouse mAb #9139 对经干扰素-α (100 ng/mL, 5”) 处理的血清饥饿 HeLa 细胞的裂解物 (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 时的单一靶标条带(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。
Immunoprecipitation Image 1: Stat3 (124H6) Mouse mAb Expand Image
对 Hela 细胞提取物的 Stat3 进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 用 Mouse (G3A1) mAb IgG1 Isotype Control #5415 进行沉淀,泳道 3 为 Stat3 (124H6) Mouse mAb #9139。
Immunohistochemistry Image 1: Stat3 (124H6) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 2: Stat3 (124H6) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 3: Stat3 (124H6) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 4: Stat3 (124H6) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human papillary carcinoma of the breast using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 1: Stat3 (124H6) Mouse mAb Expand Image
使用 Stat3 (124H6) Mouse mAb,对石蜡包埋的人乳腺癌组织(左图)以及人肺癌组织(右图)进行免疫组织化学分析,乳腺癌组织显示有细胞核和细胞质染色,肺癌组织显示细胞质染色。
Immunofluorescence Image 1: Stat3 (124H6) Mouse mAb Expand Image
对血清饥饿(左图)或 INF α 处理(右图)并用 Stat3 (124H6) Mouse mAb(绿色)标记的 HeLa 细胞,进行共聚焦免疫荧光分析。
Flow Cytometry Image 1: Stat3 (124H6) Mouse mAb Expand Image
使用 Stat3 (124H6) Mouse mAb(实线)或浓度匹配的 Mouse (G3A1) mAb IgG1 Isotype Control #5415(虚线)对 PC-3 细胞(蓝色)和 HeLa 细胞(绿色)进行流式细胞分析。使用 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 作为二抗。
Chromatin Immunoprecipitation Image 1: Stat3 (124H6) Mouse mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对饥饿过夜后用 IL-6(100 ng/ml)处理 30 分钟的 Hep G2 细胞中提取的交联染色质,在加入 Stat3 (124H6) Mouse mAb 或 Normal Rabbit IgG #2729 后,进行染色质免疫沉淀。使用 human IRF-1 promoter primers、SimpleChIP® Human c-Fos Promoter Primers #4663 和 SimpleChIP® Human α Satellite Repeat Primers #4486 并通过实时 PCR 对富集的 DNA 进行定量分析。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。
CUT and RUN Image 1: Stat3 (124H6) Mouse mAb Expand Image
使用 CUT&RUN Assay Kit #86652 ,以饥饿过夜且经 IL-6(100 ng/ml) 处理 30 分钟的 Hep G2 细胞和 Stat3 (124H6) Mouse mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 文库。该图显示在 Stat3 的已知靶标基因 BRD8 基因内的结合(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: Stat3 (124H6) Mouse mAb Expand Image
使用 CUT&RUN Assay Kit #86652 ,以饥饿过夜且经 IL-6 (100 ng/ml) 处理 30 分钟的 Hep G2 细胞和 Stat3 (124H6) Mouse mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN。使用人 BRD8 启动子引物和人 PAIS3 上游引物,通过实时 PCR 对富集的 DNA 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。
CUT and RUN Image 3: Stat3 (124H6) Mouse mAb Expand Image
使用 CUT&RUN Assay Kit #86652 ,以饥饿过夜且经 IL-6(100 ng/ml) 处理 30 分钟的 Hep G2 细胞和 Stat3 (124H6) Mouse mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示在染色体 5(上图)内的结合,包括在 Stat3 的已知靶标基因 BRD8 基因(下图)内(参见包含 CUT&RUN-qPCR 数据的其他结果图)。