Revision 1
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, IP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

75

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q96CV9

Entrez-Gene Id:

10133

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 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.

Specificity / Sensitivity

Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb recognizes endogenous levels of optineurin protein only when phosphorylated at Ser177.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser177 of human optineurin protein.

Background

Optineurin is a signaling protein involved in maintenance of the Golgi complex, membrane trafficking, NF-κB, and interferon signaling. Mutations in the gene encoding optineurin have been associated with human diseases including glaucoma, Paget disease of bone, and amyotrophic lateral sclerosis (ALS) (1-2). Optineurin is thought to contribute to these pathologies through regulation of inflammatory signaling, autophagy, and mitophagy (1, 3). The NF-κB-activating kinase/TANK-binding kinase 1 (NAK/TBK1) phosphorylates optineurin at serine 177, regulating optineurin’s role in autophagy and mitophagy (4-6). The tumor suppressor HACE1 ubiquitylates optineurin, promoting the interaction of optineurin with the autophagy receptor p62/SQSTM1 (7).
Phosphorylation of optineurin at serine 177 by TBK1 enhances binding to LC3 and promotes autophagic clearance (8). Additional studies also suggest that serine 177 is phosphorylated during mitosis by PLK1 (9). In addition to serine 177, TBK1 also phosphorylates optineurin at serine 473 and 513, which can enhance its binding to ubiquitin chains and promote mitophagy (5,6).

  1. Ying, H. and Yue, B.Y. (2016) Exp Eye Res 144, 73-80.
  2. Slowicka, K. et al. (2016) Trends Immunol 37, 621-33.
  3. Markovinovic, A. et al. (2017) Prog Neurobiol 154, 1-20.
  4. Moore, A.S. and Holzbaur, E.L. (2016) Proc Natl Acad Sci U S A 113, E3349-58.
  5. Richter, B. et al. (2016) Proc Natl Acad Sci U S A 113, 4039-44.
  6. Heo, J.M. et al. (2015) Mol Cell 60, 7-20.
  7. Liu, Z. et al. (2014) Cancer Cell 26, 106-20.
  8. Wild, P. et al. (2011) Science 333, 228-33.
  9. Kachaner, D. et al. (2012) Mol Cell 45, 553-66.

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 IP: Immunoprecipitation

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.
XP is a registered 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 1
#31304

Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb

Western Blotting Image 1: Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb Expand Image
使用 Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染空载 (-) 或转染表达带 Myc/DDK 标签的人视神经蛋白 (hOptineurin-Myc/DDK; +) 和带 GST 标签的人 IKK-ε (hIKK-ε-GST; +)(如图所示)的表达载体的 293T 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 2: Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb Expand Image
使用 Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图),对未经处理 (-) 或已经过羰基氰化物-3-氯苯腙(CCCP;100 μM,2小时; +)处理的 C2C12 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 3: Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb Expand Image
使用 Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb(上图)或 Optineurin (D2L8S) Rabbit mAb #58981(下图)对经过羰基氰酯-3-氯苯基腙(CCCP;100 μM,2 小时)处理的 HDLM-2 细胞的提取物以及未经 (-) 或已经 λ 磷酸酶和牛小肠磷酸酶 (λPPase/CIP; +) 处理的裂解物进行蛋白质印迹分析。
Western Blotting Image 4: Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb Expand Image
使用 Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb(上图)、Optineurin (D2L8S) Rabbit mAb #58981(中图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对未经 (-) 或已经过羰基氰酯-3-氯苯基腙(CCCP;100 μM,2 小时;+)和/或 BX-795(1 μM, 2 小时;+)处理(如图所示)的 Capan-2 细胞提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb Expand Image
对用羰基氰酯-3-氯苯基腙处理(CCCP;100 μM,2 小时)的 HDLM-2 细胞的 phospho-optineurin (Ser177) protein 进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb。使用 Phospho-Optineurin (Ser177) (E8L9I) Rabbit mAb 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。