Revision 3

#85493Store at -20C

1 Kit

(9 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

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Web: [email protected] cellsignal.com

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-Atg14 (Ser29) (D4B8M) Rabbit mAb 92340 20 µl 65 kDa Rabbit IgG
Atg14 (D1A1N) Rabbit mAb 96752 20 µl 65 kDa Rabbit IgG
Phospho-Beclin-1 (Ser15) (D4B7R) Rabbit mAb 84966 20 µl 60 kDa Rabbit IgG
Beclin-1 (D40C5) Rabbit mAb 3495 20 µl 60 kDa Rabbit IgG
Phospho-Atg13 (Ser355) (D6J1W) Rabbit mAb 26839 20 µl 72 kDa Rabbit IgG
Atg13 (D4P1K) Rabbit mAb 13273 20 µl 72 kDa Rabbit IgG
ULK1 (D8H5) Rabbit mAb 8054 20 µl 150 kDa Rabbit IgG
Phospho-ULK1 (Ser757) (D7O6U) Rabbit mAb 14202 20 µl 140-150 kDa Rabbit IgG
Phospho-ULK1 (Ser555) (D1H4) Rabbit mAb 5869 20 µl 140-150 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The ULK1 Substrate Antibody Sampler Kit provides an economical means of detecting the activity of ULK1 using phospho-specific and control antibodies. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

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.

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 is an inhibitor of autophagy, phosphorylates ULK1 at Ser757 and disrupts the interaction between ULK1 and AMPK (17). ULK1 has been shown to phoshorylate several targets in the autophagy pathway, including Ser29 of Atg14, Ser15 of Beclin-1, and Ser355 of Atg13 (19-22).

  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.
  19. Park, J.M. et al. (2016) Autophagy 12, 547-64.
  20. Russell, R.C. et al. (2013) Nat Cell Biol 15, 741-50.
  21. Joo, J.H. et al. (2011) Mol Cell 43, 572-85.
  22. Egan, D.F. et al. (2015) Mol Cell 59, 285-97.

Background References

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