Revision 3

#28999Store 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, IHC-P, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

160

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13464

Entrez-Gene Id:

6093

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600

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.

Specificity / Sensitivity

ROCK1 (E2G4N) Rabbit mAb recognizes endogenous levels of total ROCK1 protein. Non-specific staining was observed in placenta by immunohistochemistry. In some cell lines, this antibody cross-reacts with 45-55 kDa bands of unknown origin by western blot.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the central region of human ROCK1 protein.

Background

ROCK (Rho-associated kinase), a family of serine/threonine kinases, is an important downstream target of Rho-GTPase and plays an important role in Rho-mediated signaling. Two isoforms of ROCK have been identified: ROCK1 and ROCK2. ROCK is composed of N-terminal catalytic, coiled-coil, and C-terminal PH (pleckstrin homology) domains. The C-terminus of ROCK negatively regulates its kinase activity (1,2). ROCK1 is cleaved by caspase-3 at a conserved DETD1113/G sequence resulting in loss of its C-terminal inhibitory domain (3). ROCK2 is directly cleaved by granzyme B (grB). Cleavage activates ROCK and leads to phosphorylation of myosin light chain (MLC) and inhibition of myosin phosphatase (4). This phosphorylation may account for the mechanism by which Rho regulates cytokinesis, cell motility, cell membrane blebbing during apoptosis, and smooth muscle contraction (5-7).

  1. Nakagawa, O. et al. (1996) FEBS Lett. 392, 189-193.
  2. Lee, J.H. et al. (2004) J. Cell. Biol. 167, 327-337.
  3. Sebbagh, M. et al. (2001) Nat Cell Biol 3, 346-52.
  4. Sebbagh, M. et al. (2005) J Exp Med 201, 465-71.
  5. Amano, M. et al. (1996) J Biol Chem 271, 20246-9.
  6. Kureishi, Y. et al. (1997) J Biol Chem 272, 12257-60.
  7. Totsukawa, G. et al. (2000) J Cell Biol 150, 797-806.

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™ IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

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 3
#28999

ROCK1 (E2G4N) Rabbit mAb

Western Blotting Image 1: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对来自各种细胞系的提取物进行蛋白质印迹分析。ROCK1 蛋白在 KATO III 细胞中低表达与预测的表达模式一致。
Western Blotting Image 1: ROCK1 (E2G4N) Rabbit mAb Expand Image

Simple Western™ analysis of lysates (0.1 mg/mL) from C6 cells using ROCK1 (E2G4N) Rabbit mAb #28999. 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。

Immunohistochemistry Image 1: ROCK1 (E2G4N) Rabbit mAb Expand Image

使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人食管癌组织进行免疫组织化学分析。

Immunohistochemistry Image 2: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人肝细胞癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人尿路上皮癌组织进行免疫组织化学分析。
Immunohistochemistry Image 4: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人 B 细胞非霍奇金氏淋巴瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 5: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人乳头状甲状腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 6: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人结肠癌组织进行免疫组织化学分析。
Immunohistochemistry Image 7: ROCK1 (E2G4N) Rabbit mAb Expand Image

使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人正常食道组织进行免疫组织化学分析。

Immunohistochemistry Image 8: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的人正常小肠组织进行免疫组织化学分析。
Immunohistochemistry Image 9: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的 4T1 同基因肿瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 10: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的小鼠正常睾丸组织进行免疫组织化学分析。
Immunohistochemistry Image 11: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的小鼠正常肺组织进行免疫组织化学分析。
Immunohistochemistry Image 12: ROCK1 (E2G4N) Rabbit mAb Expand Image
与浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)比较,使用 ROCK1 (E2G4N) Rabbit mAb(左图)对石蜡包埋的人正常肾脏正常进行免疫组织化学分析。
Immunohistochemistry Image 13: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb 对石蜡包埋的 SU-DHL-4 细胞沉淀物(左图,阳性)或 KATO III 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunofluorescence Image 1: ROCK1 (E2G4N) Rabbit mAb Expand Image
使用 ROCK1 (E2G4N) Rabbit mAb(绿色)和 DAPI #4083(蓝色)对 MCF7 细胞(左图,阳性)和 KATO III 细胞(右图,阴性)进行共聚焦免疫荧光分析。