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Anti-Phospho-ULK1 (S556) Rabbit Monoclonal Antibody

Immunofluorescent analysis using the Antibody at 1:50 dilution.
Immunofluorescent analysis using the Antibody at 1:150 dilution.
Figure 1. Western blot analysis of ULK1 using anti-ULK1 antibody (M00584S556).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human 293T whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ULK1 antigen affinity purified monoclonal antibody (Catalog # M00584S556) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ULK1 at approximately 150 kDa. The expected band size for ULK1 is at 113 kDa.

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Product Information
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	WB
Primary Accession	O75385
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-Phospho-ULK1 (S556) Rabbit Monoclonal Antibody . Tested in WB, ICC/IF applications. This antibody reacts with Human, Mouse.

Additional Information
Gene ID	8408
Other Names	Serine/threonine-protein kinase ULK1, 2.7.11.1, Autophagy-related protein 1 homolog, ATG1, hATG1, Unc-51-like kinase 1, ULK1 {ECO:0000303\|PubMed:9693035, ECO:0000312\|HGNC:HGNC:12558}
Calculated MW	112631 Da
Application Details	WB 1:500-1:2000 ICC/IF 1:50-1:200
Source	Eukaryota
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: 31U30
Immunogen	A synthesized peptide derived from human Phospho-ULK1 (S556)
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	ULK1 {ECO:0000303\|PubMed:9693035, ECO:0000312\|HGNC:HGNC:12558}
Function	Serine/threonine-protein kinase involved in autophagy in response to starvation (PubMed:18936157, PubMed:21460634, PubMed:21795849, PubMed:23524951, PubMed:25040165, PubMed:29487085, PubMed:31123703). Acts upstream of phosphatidylinositol 3-kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes (PubMed:18936157, PubMed:21460634, PubMed:21795849, PubMed:25040165, PubMed:39384743). 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 (PubMed:21795849). 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 (PubMed:21460634). May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences (PubMed:18936157). Plays a role early in neuronal differentiation and is required for granule cell axon formation (PubMed:11146101). Also phosphorylates SESN2 and SQSTM1 to regulate autophagy (PubMed:25040165, PubMed:37306101). Phosphorylates FLCN, promoting autophagy (PubMed:25126726). Phosphorylates AMBRA1 in response to autophagy induction, releasing AMBRA1 from the cytoskeletal docking site to induce autophagosome nucleation (PubMed:20921139). Phosphorylates ATG4B, leading to inhibit autophagy by decreasing both proteolytic activation and delipidation activities of ATG4B (PubMed:28821708).
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