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

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

68

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q676U5

Entrez-Gene Id:

55054

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

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-Atg16L1 (Ser278) (E7K6H) Rabbit mAb recognizes endogenous levels of Atg16L1 protein only when phosphorylated at Ser278. This site is present on Atg16L1β, but not Atg16L1α.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser278 of human Atg16L1 protein.

Background

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents. Control of autophagy was largely discovered in yeast and involves proteins encoded by a set of autophagy-related genes (Atg) (1). Formation of autophagic vesicles requires a pair of essential ubiquitin-like conjugation systems, Atg12-Atg5 and Atg8 (LC3)-phosphatidylethanolamine (LC3-PE), which are widely conserved in eukaryotes (2).Mammalian Atg16L1, containing an amino-terminal coiled-coil domain and carboxyl-terminal WD-repeats, has multiple isoforms produced by alternative splicing (3,4). Atg16L1 provides a functional link between the two crucial ubiquitin-like conjugation systems of autophagy. Atg16L1 binds Atg5 of the Atg12-Atg5 conjugate forming an 800 kDa multimeric complex (3). The Atg12-Atg-5-Atg16L1 complex localizes to pre-autophagosomal membranes, where it determines the site of LC3 lipidation and catalyzes the reaction required for the formation of mature autophagosomes (3,5). Genome-wide association scanning revealed variations in the Atg16L1 gene associated with Crohn's disease (6,7). Mice lacking the coiled-coil domain of Atg16L1 have impaired autophagosome formation and elevated inflammatory cytokines, consistent with its role in inflammatory disease pathogenesis (8). Hypomorphic Atg16L1 mice also show defects in autophagy and abnormalities in intestinal Paneth cell function similar to that found in Crohn's disease (9).

ULK1-mediated phosphorylation of Atg16L1 at Ser278 promotes xenophagy, a selective autophagic process directed against pathogens (10).

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
  2. Ohsumi, Y. (2001) Nat Rev Mol Cell Biol 2, 211-6.
  3. Mizushima, N. et al. (2003) J Cell Sci 116, 1679-88.
  4. Zheng, H. et al. (2004) DNA Seq 15, 303-5.
  5. Fujita, N. et al. (2008) Mol Biol Cell 19, 2092-100.
  6. Hampe, J. et al. (2007) Nat Genet 39, 207-11.
  7. Rioux, J.D. et al. (2007) Nat Genet 39, 596-604.
  8. Saitoh, T. et al. (2008) Nature 456, 264-8.
  9. Cadwell, K. et al. (2008) Nature 456, 259-63.
  10. Alsaadi, R.M. et al. (2019) EMBO Rep 20, e46885.

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

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 1
#45511

Phospho-Atg16L1 (Ser278) (E7K6H) Rabbit mAb

Western Blotting Image 1: Phospho-Atg16L1 (Ser278) (E7K6H) Rabbit mAb Expand Image
使用 Phospho-Atg16L1 (Ser278) (E7K6H) Rabbit mAb(上图)、Atg16L1 (D6D5) Rabbit mAb #8089(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或经 Torin 1 #14379 处理(250 nM,2 小时;+)的 C2C12 细胞和 H-4-II-E 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 2: Phospho-Atg16L1 (Ser278) (E7K6H) Rabbit mAb Expand Image
使用 Phospho-Atg16L1 (Ser278) (E7K6H) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对空载转染 (-) 或经表达 Myc/DDK 标记的全长人 Atg16L1 蛋白(hAtg16L1-Myc/DDK;+)和/或小鼠 ULK1 蛋白(mULK1;+)的构建体转染的 293T 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 3: Phospho-Atg16L1 (Ser278) (E7K6H) Rabbit mAb Expand Image
使用 Phospho-Atg16L1 (Ser278) (E7K6H) Rabbit mAb(上图)、Atg16L1 (D6D5) Rabbit mAb #8089(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或经 Torin 1 #14379 处理(250 nM,2 小时;+)和/或经 ULK1 抑制剂 SBI-0206965 #29089 处理(50 μM,2 小时;+)的 AsPC-1 细胞、Calu-1 细胞和 HCT 116 细胞的提取物进行蛋白质印迹分析。