Phospho-ULK1 (Ser317) Antibody - #AF2301

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Product: Phospho-ULK1 (Ser317) Antibody
Catalog: AF2301
Description: Rabbit polyclonal antibody to Phospho-ULK1 (Ser317)
Application: WB IHC IF/ICC
Cited expt.: WB
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 120~150kD; 113kD(Calculated).
Uniprot: O75385
RRID: AB_2845315

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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, IF/ICC 1:100-1:500
*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(90%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-ULK (Ser317) Antibody detects endogenous levels of ULK1 only when phosphorylated at Ser317.
RRID:
AB_2845315
Cite Format: Affinity Biosciences Cat# AF2301, RRID:AB_2845315.
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 ULK around the phosphorylation site of Ser317.

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
Xenopus
100
Rabbit
100
Chicken
90
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). Prototheca bovis induces autophagy in bovine mammary epithelial cells via the HIF-1α and AMPKα/ULK1 pathway. Frontiers in Immunology, 2022 (PubMed: 36148236) [IF=5.7]

Application: WB    Species: bovine    Sample: bMECs

Figure 2 P. bovis infection induced autophagy via activated AMPKα/ULK1 signaling in bMECs. (A) and (B) Western blot analyses of AMPKα, p-AMPKα, p-mTOR, ULK1, p-ULK1, SQSTM1/p62, Atg5 and LC3II/LC3I in bMECs. The right panel is protein quantification with ImageJ software. The bMECs were infected with P. bovis and treated with rapamycin (20 µM) for 6 hAdditionally, bMECs were pretreated with 3MA (5 mM) for 2 h, and then infected with P. bovis for 6 h Data represent means ± SD of 3 independent experiments, compared to the control group, *P < 0.05 and **P < 0.01; compared to the P. bovis group
2). Artesunate reverses LPS tolerance by promoting ULK1-mediated autophagy through interference with the CaMKII-IP3R-CaMKKβ pathway. International Immunopharmacology, 2020 (PubMed: 32759048) [IF=4.8]

Application: WB    Species: mouse    Sample: PMs

Fig. 5. |Artesunate suppressed the excessive phosphorylation of AMPKα topromote ULK1 activation and reverse LPS tolerance.) (E-F) Tolerant PMs were stimulated with 100 ng/mL LPS alone or with Compound C (2–30 μM) for 2 h, bafilomycin A1 (20 nM)was used to evaluate the effect on autophagic flow. Phosphorylation of mTOR, AMPKα, LC3-II, and LC3-I (E) orphosphorylation sites of Ser317, 637, and 757 in ULK1 (F) were determined by western blot analysis. Both the representative blots and the densitometry data (n = 3) were shown.
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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