Revision 3

#37762Store 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

SENSITIVITY:

Endogenous

MW (kDa):

140

SOURCE:

Rabbit

UniProt ID:

#O75385

Entrez-Gene Id:

8408

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 and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-ULK1 (Ser317) Antibody recognizes endogenous levels of ULK1 protein only when phosphorylated at Ser317.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser317 of human ULK1 protein. Antibodies are purified by peptide affinity chromatography.

Background

Two related serine/threonine kinases, UNC-51-like kinase 1 and 2 (ULK1, ULK2), were discovered as mammalian homologs of the C. elegans gene unc-51 in which mutants exhibited abnormal axonal extension and growth (1-4). Both proteins are widely expressed and contain an amino-terminal kinase domain followed by a central proline/serine rich domain and a highly conserved carboxy-terminal domain. The roles of ULK1 and ULK2 in axon growth have been linked to studies showing that the kinases are localized to neuronal growth cones and are involved in endocytosis of critical growth factors, such as NGF (5). Yeast two-hybrid studies found ULK1/2 associated with modulators of the endocytic pathway, SynGAP, and syntenin (6). Structural similarity of ULK1/2 has also been recognized with the yeast autophagy protein Atg1/Apg1 (7). Knockdown experiments using siRNA demonstrated that ULK1 is essential for autophagy (8), a catabolic process for the degradation of bulk cytoplasmic contents (9,10). It appears that Atg1/ULK1 can act as a convergence point for multiple signals that control autophagy (11), and can bind to several autophagy-related (Atg) proteins, regulating phosphorylation states and protein trafficking (12-16).~AMPK, activated during low nutrient conditions, directly phosphorylates ULK1 at multiple sites, including Ser317, Ser555, and Ser777 (17,18). Conversely, mTOR, which is a regulator of cell growth and an inhibitor of autophagy, phosphorylates ULK1 at Ser757 and disrupts the interaction between ULK1 and AMPK (17).

  1. Ogura, K. et al. (1994) Genes Dev 8, 2389-400.
  2. Kuroyanagi, H. et al. (1998) Genomics 51, 76-85.
  3. Yan, J. et al. (1998) Biochem Biophys Res Commun 246, 222-7.
  4. Yan, J. et al. (1999) Oncogene 18, 5850-9.
  5. Zhou, X. et al. (2007) Proc Natl Acad Sci USA 104, 5842-7.
  6. Tomoda, T. et al. (2004) Genes Dev 18, 541-58.
  7. Matsuura, A. et al. (1997) Gene 192, 245-50.
  8. Chan, E.Y. et al. (2007) J Biol Chem 282, 25464-74.
  9. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
  10. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
  11. Stephan, J.S. and Herman, P.K. (2006) Autophagy 2, 146-8.
  12. Okazaki, N. et al. (2000) Brain Res Mol Brain Res 85, 1-12.
  13. Young, A.R. et al. (2006) J Cell Sci 119, 3888-900.
  14. Kamada, Y. et al. (2000) J Cell Biol 150, 1507-13.
  15. Lee, S.B. et al. (2007) EMBO Rep 8, 360-5.
  16. Hara, T. et al. (2008) J Cell Biol 181, 497-510.
  17. Kim, J. et al. (2011) Nat Cell Biol 13, 132-41.
  18. Egan, D.F. et al. (2011) Science 331, 456-61.

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 BSA, 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.

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Revision 3
#37762

Phospho-ULK1 (Ser317) Antibody

Western Blotting Image 1: Phospho-ULK1 (Ser317) Antibody Expand Image
使用 Phospho-ULK1 (Ser317) Antibody(上图)、ULK1 (D8H5) Rabbit mAb #8054(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对经 AMPK 激活化合物991处理(50 μM,1 小时)以及随后未处理 (-) 或经 Lambda-Phosphatase (λ-Phosphatase) 和 Calf Intestinal Phosphatase (CIP) 处理 (+) 的 PANC-1 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 2: Phospho-ULK1 (Ser317) Antibody Expand Image
使用 Phospho-ULK1 (Ser317) Antibody(上图)、ULK1 (D8H5) Rabbit mAb #8054(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对未经处理 (-) 或经 AMPK 激活化合物991(50 μM,1 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-ULK1 (Ser317) Antibody Expand Image
使用 Phospho-ULK1 (Ser317) Antibody(上图)、ULK1 (D8H5) Rabbit mAb #8054(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对未经处理 (-) 或经 AMPK 激活化合物991(50 μM,1 小时)处理的 PANC-1 细胞提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Phospho-ULK1 (Ser317) Antibody Expand Image
对经 AMPK 激活化合物处理(50 μM,1 小时)的 PANC-1 细胞提取物的磷酸化 ULK1 (Ser317) 蛋白进行免疫沉淀 991 。泳道 1 为 10% input,泳道 2 为 Normal Rabbit IgG #2729,泳道 3 为 Phospho-ULK1 (Ser317) Antibody。使用 Phospho-ULK1 (Ser317) Antibody 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。