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Phospho-ULK1(S556) Antibody

Peptide Affinity Purified Rabbit Polyclonal Antibody (Pab)

     
  • 3 - Phospho-ULK1(S556) Antibody AP3804a
    Fluorescent image of U251 cells stained with ULK1 (phospho S556) antibody. U251 cells were treated with Chloroquine (50 μM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP3804a ULK1 (phospho S556) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). ULK1 (phospho S556) immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.
  • 6 - Phospho-ULK1(S556) Antibody AP3804a
    Dot blot analysis of ULK1 Antibody (Phospho S556) Phospho-specific Pab (Cat. #AP3804a) on nitrocellulose membrane. 50ng of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.6ug per ml.
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Product info
Application
  • Applications Legend:
  • E=ELISA
  • WB=Western Blotting
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin)
  • IP=Immunoprecipitation
  • IF=Immunofluorescence
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • FC=Flow Cytometry
  • DB=Dot Blot
IF, DB, E
Primary Accession O75385
Other Accession NP_003556.1
Reactivity Human
Host Rabbit
Clonality Polyclonal
Isotype Rabbit Ig
Calculated MW 112631 Da
Additional info
Gene ID 8408
Other Names Serine/threonine-protein kinase ULK1, Autophagy-related protein 1 homolog, ATG1, hATG1, Unc-51-like kinase 1, ULK1, KIAA0722
Target/Specificity This ULK1 Antibody is generated from rabbits immunized with a KLH conjugated synthetic phosphopeptide corresponding to amino acid residues surrounding S556 of human ULK1.
Dilution IF~~1:200
DB~~1:500
Format Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by two-step phosphospecific peptide affinity purification.
StorageMaintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
PrecautionsPhospho-ULK1(S556) Antibody is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name ULK1
Synonyms KIAA0722
Function Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3- kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2 and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. May also phosphorylate SESN2 and SQSTM1 to regulate autophagy (PubMed:25040165).
Cellular Location Cytoplasm, cytosol. Preautophagosomal structure. Note=Under starvation conditions, is localized to puncate structures primarily representing the isolation membrane that sequesters a portion of the cytoplasm resulting in the formation of an autophagosome
Tissue Location Ubiquitously expressed. Detected in the following adult tissues: skeletal muscle, heart, pancreas, brain, placenta, liver, kidney, and lung
Research Areas

BACKGROUND

Involved in autophagy. Required for autophagosome formation (By similarity). Target of the TOR kinase signaling pathway that regulates autophagy through the control of phosphorylation status of ATG13/KIAA0652 and ULK1, and the regulation of the ATG13-ULK1-RB1CC1 complex (By similarity). Phosphorylates ATG13/KIAA0652. Involved in axon growth (By similarity). Plays an essential role in neurite extension of cerebellar granule cells (By similarity).

REFERENCES

References for protein:
1.Mercer, C.A., et al. Autophagy 5(5):649-662(2009)
2.Ganley, I.G., et al. J. Biol. Chem. 284(18):12297-12305(2009)
3.Jung, C.H., et al. Mol. Biol. Cell 20(7):1992-2003(2009)
4.Hosokawa, N., et al. Mol. Biol. Cell 20(7):1981-1991(2009)
5.Chan, E.Y. Sci Signal 2 (84), PE51 (2009)
References for U251 cell line:
1. Westermark B.; Pontén J.; Hugosson R. (1973).” Determinants for the establishment of permanent tissue culture lines from human gliomas”. Acta Pathol Microbiol Scand A. 81:791-805. [PMID: 4359449].
2. Pontén, J.,Westermark B. (1978).” Properties of Human Malignant Glioma Cells in Vitro”. Medical Biology 56: 184-193.[PMID: 359950].
3. Geng Y.;Kohli L.; Klocke B.J.; Roth K.A.(2010). “Chloroquine-induced autophagic vacuole accumulation and cell death in glioma cells is p53 independent”. Neuro Oncol. 12(5): 473–481.[ PMID: 20406898].

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