Phospho-ULK1 (Ser555)[Ser556] Antibody - #AF7148

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Product: Phospho-ULK1 (Ser555)[Ser556] Antibody
Catalog: AF7148
Description: Rabbit polyclonal antibody to Phospho-ULK1 (Ser555)[Ser556]
Application: WB IHC
Cited expt.: WB, IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 120kDa; 113kD(Calculated).
Uniprot: O75385
RRID: AB_2843588

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(89%), Xenopus(88%)
Clonality:
Polyclonal
Specificity:
Phospho-ULK1 (Ser555) Antibody detects endogenous levels of ULK1 only when phosphorylated at Ser555 of mouse ULK1 or Ser556 of human ULK1.
RRID:
AB_2843588
Cite Format: Affinity Biosciences Cat# AF7148, RRID:AB_2843588.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

ATG 1; ATG1; ATG1 autophagy related 1 homolog; ATG1A; Autophagy related protein 1 homolog; Autophagy-related protein 1 homolog; FLJ38455; FLJ46475; hATG1; KIAA0722; Serine/threonine protein kinase ULK1; Serine/threonine protein kinase Unc51.1; Serine/threonine-protein kinase ULK1; ULK 1; ULK1; ULK1_HUMAN; Unc 51 (C. elegans) like kinase 1; UNC 51; Unc 51 like kinase 1; Unc-51 like kinase 1 (C. elegans); Unc-51-like kinase 1; UNC51; UNC51, C. elegans, homolog of; Unc51.1;

Immunogens

Immunogen:

A synthesized peptide derived from human ULK1 around the phosphorylation site of Ser556.

Uniprot:

O75385(ULK1_HUMAN) >>Visit HPA database.

Gene(ID):
ULK1(8408)
Expression:
O75385 ULK1_HUMAN:

Ubiquitously expressed. Detected in the following adult tissues: skeletal muscle, heart, pancreas, brain, placenta, liver, kidney, and lung.

Sequence:
MEPGRGGTETVGKFEFSRKDLIGHGAFAVVFKGRHREKHDLEVAVKCINKKNLAKSQTLLGKEIKILKELKHENIVALYDFQEMANSVYLVMEYCNGGDLADYLHAMRTLSEDTIRLFLQQIAGAMRLLHSKGIIHRDLKPQNILLSNPAGRRANPNSIRVKIADFGFARYLQSNMMAATLCGSPMYMAPEVIMSQHYDGKADLWSIGTIVYQCLTGKAPFQASSPQDLRLFYEKNKTLVPTIPRETSAPLRQLLLALLQRNHKDRMDFDEFFHHPFLDASPSVRKSPPVPVPSYPSSGSGSSSSSSSTSHLASPPSLGEMQQLQKTLASPADTAGFLHSSRDSGGSKDSSCDTDDFVMVPAQFPGDLVAEAPSAKPPPDSLMCSGSSLVASAGLESHGRTPSPSPPCSSSPSPSGRAGPFSSSRCGASVPIPVPTQVQNYQRIERNLQSPTQFQTPRSSAIRRSGSTSPLGFARASPSPPAHAEHGGVLARKMSLGGGRPYTPSPQVGTIPERPGWSGTPSPQGAEMRGGRSPRPGSSAPEHSPRTSGLGCRLHSAPNLSDLHVVRPKLPKPPTDPLGAVFSPPQASPPQPSHGLQSCRNLRGSPKLPDFLQRNPLPPILGSPTKAVPSFDFPKTPSSQNLLALLARQGVVMTPPRNRTLPDLSEVGPFHGQPLGPGLRPGEDPKGPFGRSFSTSRLTDLLLKAAFGTQAPDPGSTESLQEKPMEIAPSAGFGGSLHPGARAGGTSSPSPVVFTVGSPPSGSTPPQGPRTRMFSAGPTGSASSSARHLVPGPCSEAPAPELPAPGHGCSFADPITANLEGAVTFEAPDLPEETLMEQEHTEILRGLRFTLLFVQHVLEIAALKGSASEAAGGPEYQLQESVVADQISLLSREWGFAEQLVLYLKVAELLSSGLQSAIDQIRAGKLCLSSTVKQVVRRLNELYKASVVSCQGLSLRLQRFFLDKQRLLDRIHSITAERLIFSHAVQMVQSAALDEMFQHREGCVPRYHKALLLLEGLQHMLSDQADIENVTKCKLCIERRLSALLTGICA

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Chicken
89
Xenopus
88
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

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.

PTMs:

Autophosphorylated. Phosphorylated under nutrient-rich conditions; dephosphorylated during starvation or following treatment with rapamycin. Under nutrient sufficiency, phosphorylated by MTOR/mTOR, disrupting the interaction with AMPK and preventing activation of ULK1 (By similarity). In response to nutrient limitation, phosphorylated and activated by AMPK, leading to activate autophagy.

Acetylated by KAT5/TIP60 under autophagy induction, promoting protein kinase activity.

Subcellular 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.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Ubiquitously expressed. Detected in the following adult tissues: skeletal muscle, heart, pancreas, brain, placenta, liver, kidney, and lung.

Family&Domains:

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. APG1/unc-51/ULK1 subfamily.

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway.   (View pathway)

References

1). SARS-CoV-2 ORF3a induces RETREG1/FAM134B-dependent reticulophagy and triggers sequential ER stress and inflammatory responses during SARS-CoV-2 infection. Autophagy, 2022 (PubMed: 35239449) [IF=14.6]
2). Vasorin Deletion in C57BL/6J Mice Induces Hepatocyte Autophagy through Glycogen-Mediated mTOR Regulation. Nutrients, 2022 (PubMed: 36079859) [IF=5.9]

Application: WB    Species: Mice    Sample: liver

Figure 6. Vasn deletion induced autophagy by regulating the mTOR-ULK1 signaling pathway. (A) Pathway enrichment results of target genes of differential expression microRNAs, where the mTOR pathway is of the Top 1 that has been significantly enriched (red highlightened). (B) Immunohistochemical analysis of mTOR signaling protein expression in the liver of WT, Vasn+/−, and Vasn−/− mice; bar = 100 μm. (C) Quantitative integrated optical density (IOD) analysis of immunohistochemical staining from ten mice per group, and three photos per sample. ** p < 0.01, *** p < 0.001. (D) Vasn deletion significantly decreased the expression of phosphorylated mTOR (S2448) and increased expression of the phosphorylated ULK1 (S555), confirmed by Western blot analysis.

Application: IHC    Species: Mice    Sample: liver

Figure 6. Vasn deletion induced autophagy by regulating the mTOR-ULK1 signaling pathway. (A) Pathway enrichment results of target genes of differential expression microRNAs, where the mTOR pathway is of the Top 1 that has been significantly enriched (red highlightened). (B) Immunohistochemical analysis of mTOR signaling protein expression in the liver of WT, Vasn+/−, and Vasn−/− mice; bar = 100 μm. (C) Quantitative integrated optical density (IOD) analysis of immunohistochemical staining from ten mice per group, and three photos per sample. ** p < 0.01, *** p < 0.001. (D) Vasn deletion significantly decreased the expression of phosphorylated mTOR (S2448) and increased expression of the phosphorylated ULK1 (S555), confirmed by Western blot analysis.
3). Xiaojianzhong decoction attenuates aspirin-induced gastric mucosal injury via the PI3K/AKT/mTOR/ULK1 and AMPK/ULK1 pathways. Pharmaceutical biology, 2023 (PubMed: 37602379) [IF=3.9]

Application: WB    Species: Mouse    Sample: stomach tissue

Figure 7. XJZD inhibits the AMPK/ULK1(ser317) pathway. (A) The expression of ULK1 was detected by immunofluorescence assay, magnification ×400, scale bar: 20 µm (n = 6). (B) Using ImageJ software to analyse the fluorescence intensity of ULK1. (C) Representative Western blot bands of AMPK and p-AMPK. (D) Protein levels of p-AMPK/AMPK. (E) The mRNA levels of ULK1. (F) Representative Western blot bands of ULK1, p-ULK1(ser555) and p-ULK1(ser317). (G, H) Protein levels of p-ULK1(ser555)/ULK1 and p-ULK1(ser317)/ULK1 (n = 3). These data are means ± SD, ##p < 0.01 and ###p < 0.001 when compared with the control group. *p < 0.05, **p < 0.01 and ***p < 0.001 when compared with the model group. ns: no significant difference.

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