| 货号 | 8359T |
| 描述 | The ULK1 Antibody Sampler Kit provides an economical way to investigate ULK1 signaling. The kit contains enough primary antibody to perform four western blots with each primary antibody. |
| 目标/特异性 | ULK1 (R600) Antibody, Raptor (24C12) Rabbit mAb, and AMPKα (D63G4) Rabbit mAb recognize total endogenous levels of the corresponding target proteins irrespective of phosphorylation state. Phospho-ULK1 (Ser555) (D1H4) Rabbit mAb and Phospho-ULK1 (Ser757) Antibody recognize endogenous levels of ULK1 only when phosphorylated at the indicated residues. Phospho-Raptor (Ser792) Antibody recognizes endogenous levels of raptor only when phosphorylated at Ser792. Phospho-AMPKα (Thr172) (40H9) Rabbit mAb recognizes endogenous levels of AMPKα only when phosphorylated at Thr172. |
| 供应商 | CST |
| 背景 | 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).Raptor mediates the binding of mTORC1 to ULK1, which phosphorylates and inhibits ULK1 under nutrient rich conditions. AMPK also associates directly with ULK1 and, upon nutrient deprivation, can readily reverse the inhibitory effect of mTORC1 by phosphorylating raptor and initiating autophagy (17,18). |
| 存放说明 | -20C |
| 参考文献 | 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. Shang, L. et al. (2011) Proc Natl Acad Sci U S A 108, 4788-93. Lee, J.W. et al. (2010) PLoS One 5, e15394. |
Western blot analysis of extracts from C2C12 cells, untreated (-) or treated (+) with Oligomycin #9996 (0.5 µM), using Phospho-AMPKα (Thr172) (40H9) Rabbit mAb #2535 (upper) or AMPKα Antibody #2532 (lower). Western blot 分析C2C12细胞的细胞提取物，未处理（-）或 Oligomycin #9996 (0.5 µM)处理（+），使用抗体是Phospho-AMPKα (Thr172) (40H9) Rabbit mAb 兔单抗#2535 (上图) 或 AMPKα Antibody #2532 (下图)。 | |
Western blot analysis of C2C12 or 293 cells, untreated (-) or treated (+) with AICAR (0.5 mM, 30 min) or Oligomycin #9996 (0.5 μM, 30 min), using Phospho-Raptor (Ser792) Antibody #2083 (upper and lower left) or Raptor (24C12) Rabbit mAb #2280 (upper and lower right). *Cross-reacting bands at 200 kDa. Western blot 分析C2C12或293细胞的细胞提取物，未处理（-）或 AICAR (0.5 mM, 30 min) 或Oligomycin #9996 (0.5 μM, 30 min)处理（+），使用抗体是Phospho-Raptor (Ser792) Antibody #2083 (上图和下图左)或Raptor (24C12) Rabbit mAb 兔单抗#2280 (上图和下图右)。 | |
Western blot analysis of extracts from various cell lines using AMPKα (D63G4) Rabbit mAb #5832. Western blot 分析多种细胞系的细胞提取物，使用抗体是AMPKα (D63G4) Rabbit mAb 兔单抗#5832。 | |
Western blot analysis of extracts from various cell lines using Raptor (24C12) Rabbit mAb #2280. Western blot分析多种细胞系的细胞提取物，使用抗体是Raptor (24C12) Rabbit mAb 兔单抗#2280。 | |
Western blot analysis of extracts from A-431 cells, untreated (-) or treated (+) with Human Epidermal Growth Factor (hEGF) #8916 (100 ng/ml, 30 min) using Phospho-ULK1 (Ser757) Antibody #6888 (upper), or α-Tubulin (11H10) Rabbit mAb #2125 (lower). Western blot检测A-431细胞提取物，未处理（-）或Human Epidermal Growth Factor (hEGF) #8916 (100 ng/ml, 30 min)处理（+），使用抗体是Phospho-ULK1 (Ser757) Antibody #6888 (上图)或α-Tubulin (11H10) Rabbit mAb 兔单抗#2125 (下图)。 | |
Western blot analysis of extracts from MCF7 cells, untreated (-) or treated (+) with Oligomycin #9996 (0.5 μM, 30 min), and C2C12 cells, untreated (-) or treated (+) with hydrogen peroxide (10 mM, 5 min), using Phospho-ULK1 (Ser555) (D1H4) Rabbit mAb #5869. Western blot 检测MCF7细胞的提取物，未处理（-）或经 Oligomycin #9996 处理（+）（0.5μM，30分钟）；C2C12细胞提取物，使用未处理（-）或过氧化氢 (10 mM, 5 min)处理（+），使用抗体为Phospho-ULK1 (Ser555) (D1H4) Rabbit mAb 兔单抗#5869。 | |
Immunohistochemical analysis of paraffin-embedded NCI-H228 cell pellets, control (left) or phenformin-treated (right), using Phospho-AMPKalpha (T172) (40H9) Rabbit mAb. | |
Western blot analysis of extracts from HeLa, K-562, C6, and Neuro-2a cells using AMPKα (D63G4) Rabbit mAb. | |
Western blot analysis of extracts from MCF7 cells, untreated or treated with oligomycin #9996 (0.5 μM, 30 minutes), and C2C12 cells, untreated or treated with hydrogen peroxide (10 mM, 5 minutes), using Phospho-ULK1 (Ser555) (D1H4) Rabbit mAb. | |
Western blot analysis of extracts from MCF7 cells, untreated or treated with oligomycin #9996 (0.5 µM, 30 minutes), using Phosho-ULK1 (Ser555) (D1H4) Rabbit mAb (left). Phoshpo-specificty is demonstrated by pre-incubating the antibody with phosphorylated (middle) or non-phoshporylated peptides (right) against a region surrounding Ser555 of ULK1. | |
Western blot analysis of extracts from A-431 cells, untreated or treated with Human Epidermal Growth Factor (hEGF) #8916 (100 ng/ml, 30 min) using Phospho-ULK1 (Ser757) Antibody (upper), or α-Tubulin (11H10) Rabbit mAb #2125 (lower). | |
Western blot analysis of extracts from RD and SH-SY5Y cells using ULK1 (R600) Antibody #4773. 使用ULK1 (R600) Antibody #4773 对RD和SH-SY5Y的细胞提取物进行 Western blot 检测。 | |
*Cross-reacting bands at 200 kDa. |
