Revision 3
#93778
Store at +4C
PathScan® RP Phospho-ULK1 (Ser757) Sandwich ELISA Kit
1 Kit
(96 assays)
Species Cross Reactivity:
H M
UniProt ID:
#O75385
Entrez-Gene Id:
#8408
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes | Product # | Quantity | Color | Storage Temp |
---|---|---|---|---|
Phospho-ULK1 (Ser757) Rabbit mAb Coated Microwells | 68745 | 96 tests | +4C | |
ULK1 Rabbit Detection mAb | 53531 | 1 ea | Red (Lyophilized) | +4C |
HRP Diluent | 13515 | 5.5 ml | Red | +4C |
TMB Substrate | 7004 | 11 ml | +4C | |
STOP Solution | 7002 | 11 ml | +4C | |
Sealing Tape | 54503 | 2 ea | +4C | |
ELISA Wash Buffer (20X) | 9801 | 25 ml | +4C | |
Cell Lysis Buffer (10X) | 9803 | 15 ml | -20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The rapid protocol (RP) PathScan® RP Phospho-ULK1 (Ser757) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of ULK1 protein phosphorylated at Ser757 in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with phospho-ULK1 (Ser757) in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of phospho-ULK1 (Ser757). Learn more about all of your ELISA kit options here.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
The PathScan® RP Phospho-ULK1 (Ser757) Sandwich ELISA Kit detects endogenous levels of ULK1 protein phosphorylated at Ser757. The kit sensitivity is shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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).
Background References
- Ogura, K. et al. (1994) Genes Dev 8, 2389-400.
- Kuroyanagi, H. et al. (1998) Genomics 51, 76-85.
- Yan, J. et al. (1998) Biochem Biophys Res Commun 246, 222-7.
- Yan, J. et al. (1999) Oncogene 18, 5850-9.
- Zhou, X. et al. (2007) Proc Natl Acad Sci USA 104, 5842-7.
- Tomoda, T. et al. (2004) Genes Dev 18, 541-58.
- Matsuura, A. et al. (1997) Gene 192, 245-50.
- Chan, E.Y. et al. (2007) J Biol Chem 282, 25464-74.
- Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
- Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
- Stephan, J.S. and Herman, P.K. (2006) Autophagy 2, 146-8.
- Okazaki, N. et al. (2000) Brain Res Mol Brain Res 85, 1-12.
- Young, A.R. et al. (2006) J Cell Sci 119, 3888-900.
- Kamada, Y. et al. (2000) J Cell Biol 150, 1507-13.
- Lee, S.B. et al. (2007) EMBO Rep 8, 360-5.
- Hara, T. et al. (2008) J Cell Biol 181, 497-510.
- Kim, J. et al. (2011) Nat Cell Biol 13, 132-41.
- Egan, D.F. et al. (2011) Science 331, 456-61.
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 G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan 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.
Limited Uses
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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 3
Figure 1. Treatment of A-172 cells with the mTOR inhibitor Torin 1 inhibits phosphorylation of ULK1 at Ser757. The relationship between lysate protein concentration from untreated and Torin 1-treated A-172 cells and the absorbance at 450 nm using the PathScan® RP Phospho-ULK1 (Ser757) Sandwich ELISA Kit #93778 is shown in the upper figure. The corresponding western blots using phospho-ULK1 (Ser757) antibody (left panel) and ULK1 antibody (right panel) are shown in the lower figure. A-172 cells were treated with Torin 1 (250 nM) for 5 hours at 37°C and then lysed.