Revision 3

#13509Store 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, IP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

100-110

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q7Z3C6

Entrez-Gene Id:

79065

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 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.

Specificity / Sensitivity

Atg9A (D4O9D) Rabbit mAb recognizes endogenous levels of total Atg9A protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly780 of human Atg9A protein.

Background

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). It is generally activated by conditions of nutrient deprivation but is also associated with a number of physiological processes including development, differentiation, neurodegeneration, infection, and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and is directed by a number of autophagy-related (Atg) genes (4).
Atg9, one of the Atg proteins identified in yeast, is essential for autophagosome formation (5). There are two human functional orthologues based on the yeast homolog Atg9p: Atg9A, which has also been identified as Atg9L1 and mAtg9, and Atg9L2, which was first reported as nitric-oxide synthase 3 antisense (NOS3AS) (6,7). Atg9A is an integral membrane protein that is required for both the initiation and the expansion of the autophagosome (6,7). Recruitment of Atg9A to the autophagosomal membrane is dynamic and transient as Atg9A also cycles between autophagy-related structures known as omegasomes, the trans-Golgi network (TGN), and endosomes, and at no point becomes a stable component of the autophagosomal membrane (6,8). The precise regulation of Atg9A trafficking is not fully clarified, yet it is suggested to involve p38 mitogen-activated protein kinase (MAPK)-binding protein p38IP and the Beclin-1-binding protein Bif-1 (9,10).

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
  3. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
  4. Klionsky, D.J. et al. (2003) Dev Cell 5, 539-45.
  5. Noda, T. et al. (2000) J Cell Biol 148, 465-80.
  6. Young, A.R. et al. (2006) J Cell Sci 119, 3888-900.
  7. Yamada, T. et al. (2005) J Biol Chem 280, 18283-90.
  8. Orsi, A. et al. (2012) Mol Biol Cell 23, 1860-73.
  9. Webber, J.L. and Tooze, S.A. (2010) EMBO J 29, 27-40.
  10. Takahashi, Y. et al. (2011) Autophagy 7, 61-73.

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.
SignalSilence is a registered 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 3
#13509

Atg9A (D4O9D) Rabbit mAb

Western Blotting Image 1: Atg9A (D4O9D) Rabbit mAb Expand Image
使用 Atg9A (D4O9D) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
Western Blotting Image 2: Atg9A (D4O9D) Rabbit mAb Expand Image
使用 Atg9A (D4O9D) Rabbit mAb(上)或 β-Actin (D6A8) Rabbit mAb #8457(下)对经 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) 或 SignalSilence® Atg9A siRNA I #7051 (+) 转染的 RD 细胞提取物进行蛋白质印迹分析。Atg9A (D4O9D) Rabbit mAb 可证实 Atg9A 表达沉默,而 β-Actin (D6A8) Rabbit mAb 用作上样对照。
Western Blotting Image 3: Atg9A (D4O9D) Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype control #3900(泳道 2)或 Atg9A (D4O9D) Rabbit mAb(泳道 3),对来自 Hep G2 细胞提取物的 Atg9A 进行免疫沉淀。泳道 1 是 10% 输入对照。使用 Atg9A (D4O9D) Rabbit mAb 进行蛋白质印迹分析。
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