Anti-Phospho-ATG1 (Ser556) Polyclonal Antibody is a Rabbit antibody targeting Phospho-ATG1 (Ser556). Anti-Phospho-ATG1 (Ser556) Polyclonal Antibody can be used in ELISA,FCM,ICC/IF,IF,IHC-Fr,IHC-P.

IgG

Human,Mouse,Rat,Rabbit (predicted:Pig,Horse)

1. phosphopeptide
non phosphopeptide
2. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Phospho-ATG1 (Ser556) Polyclonal Antibody, Unconjugated (TMAB-01393) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Blank control: A549. Primary Antibody (green line): Rabbit Anti-Phospho-ATG1 (Ser556) antibody (TMAB-01393)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 3 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 20% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
4. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-ATG1 (Ser556)) polyclonal Antibody, Unconjugated (TMAB-01393) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 3ug/test; ELISA: 1:5000-10000

Polyclonal

KLH conjugated Synthesised phosphopeptide: human ATG1/ULK1 around the phosphorylation site of Ser556

Human

ULK1

Serine/threonine-protein kinase ULK1

Other Kinases,Neurogenesis,Mitophagy fission and fusion,Autophagy

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.