Phospho-ULK1 (Ser757)[Ser758] Antibody - #AF4387

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Product: Phospho-ULK1 (Ser757)[Ser758] Antibody
Catalog: AF4387
Description: Rabbit polyclonal antibody to Phospho-ULK1 (Ser757)[Ser758]
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus
Mol.Wt.: 120~150kDa; 113kD(Calculated).
Uniprot: O75385
RRID: AB_2844452

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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%), Xenopus(88%)
Clonality:
Polyclonal
Specificity:
Phospho-ULK1 (Ser757) Antibody detects endogenous levels of ULK1 only when phosphorylated at Ser757 of mouse ULK1 or Ser758 of human ULK1.
RRID:
AB_2844452
Cite Format: Affinity Biosciences Cat# AF4387, RRID:AB_2844452.
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:
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
Xenopus
88
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O75385 As Substrate

Site PTM Type Enzyme
K13 Ubiquitination
K46 Ubiquitination
K55 Ubiquitination
K62 Ubiquitination
S111 Phosphorylation
S131 Phosphorylation
K140 Ubiquitination
S158 Phosphorylation
K162 Acetylation
K162 Ubiquitination
T180 Phosphorylation O75385 (ULK1)
S225 Phosphorylation
K235 Ubiquitination
K237 Ubiquitination
S281 Phosphorylation
S317 Phosphorylation Q13131 (PRKAA1) , P54646 (PRKAA2)
S330 Phosphorylation
S403 Phosphorylation
S405 Phosphorylation
S409 Phosphorylation
S415 Phosphorylation
S429 Phosphorylation
S450 Phosphorylation
T456 Phosphorylation
S459 Phosphorylation
S460 Phosphorylation
S465 Phosphorylation
S467 Phosphorylation
T468 Phosphorylation
S469 Phosphorylation
S477 Phosphorylation
S479 Phosphorylation
S495 Phosphorylation
S522 Phosphorylation
R529 Methylation
S533 Phosphorylation
S538 Phosphorylation
S539 Phosphorylation
S544 Phosphorylation
S556 Phosphorylation P54646 (PRKAA2)
S561 Phosphorylation
T575 Phosphorylation
S583 Phosphorylation
S588 Phosphorylation
S593 Phosphorylation
S598 Phosphorylation
S605 Phosphorylation
K607 Acetylation
S623 Phosphorylation
T625 Phosphorylation
S630 Phosphorylation
T636 Phosphorylation
S638 Phosphorylation P42345 (MTOR) , P54646 (PRKAA2) , Q13131 (PRKAA1)
S639 Phosphorylation
T654 Phosphorylation
T660 Phosphorylation
S665 Phosphorylation
S694 Phosphorylation
T695 Phosphorylation
T699 Phosphorylation
T709 Phosphorylation
S716 Phosphorylation
T717 Phosphorylation
S719 Phosphorylation
S748 Phosphorylation
S758 Phosphorylation P42345 (MTOR)
S761 Phosphorylation
T764 Phosphorylation
S775 Phosphorylation
S781 Phosphorylation
S949 Phosphorylation
S954 Phosphorylation
S1042 Phosphorylation
T1046 Phosphorylation

PTMs - O75385 As Enzyme

Substrate Site Source
A2RUS2 (DENND3) S472 Uniprot
A2RUS2 (DENND3) S490 Uniprot
O00560 (SDCBP) S6 Uniprot
O75143 (ATG13) S355 Uniprot
O75385 (ULK1) T180 Uniprot
Q13131 (PRKAA1) S360 Uniprot
Q13131 (PRKAA1) T368 Uniprot
Q13131 (PRKAA1) S397 Uniprot
Q13131 (PRKAA1) S486 Uniprot
Q13131 (PRKAA1) T488 Uniprot
Q7Z3C6 (ATG9A) S761 Uniprot
Q86WV6 (TMEM173) S366 Uniprot
Q8N122 (RPTOR) S792 Uniprot
Q8N122 (RPTOR) S855 Uniprot
Q8N122 (RPTOR) S859 Uniprot

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.

Subunit Structure:

Interacts with GABARAP and GABARAPL2. Interacts (via C-terminus) with ATG13. Part of a complex consisting of ATG13, ATG101, ULK1 and RB1CC1. Associates with the mammalian target of rapamycin complex 1 (mTORC1) through an interaction with RPTOR; the association depends on nutrient conditions and is reduced during starvation. Interacts with FEZ1; SCOC interferes with FEZ1-binding. Interacts with TBC1D14. Interacts (phosphorylated form) with TRIM5. When phosphorylated at Ser-317, interacts with MEFV and BECN1 simultaneously. Interacts with TRIM21 and IRF3, in the presence of TRIM21. Interacts with SESN2. Interacts with SQSTM1. Interacts with C9orf72. Interacts with WDR45.

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). FGF21 ameliorates septic liver injury by restraining proinflammatory macrophages activation through the autophagy/HIF-1α axis. Journal of advanced research, 2024 (PubMed: 38599281) [IF=10.7]

Application: WB    Species: Mouse    Sample:

Fig. 7. rhFGF21 promotes autophagic degradation of HIF-1α. (A, B) Representative immunofluorescence and quantification of colocalization of HIF-1α and Lamp1 in BMDMs from indicated group (n = 5). DAPI (blue), HIF-1α (red), and Lamp1 (green). Scale bars: 20 μm. (C) The expression of HIF-1α in lysates of BMDMs in indicated groups, and normalized to β-actin (n = 5). Bafilomycin A1 was added for the last 6 h. (D) BMDMs were treated as indicated, and the cell lysates were subjected to immunoprecipitation with HIF-1α or p62 antibody, followed by immunoblotting with the HIF-1α or p62 antibody. MG132 (10 μM) and Bafilomycin A1 (100 nM) were added for the last 6 h. (E) The expression of HIF-1α in lysates of BMDMs in indicated groups, and normalized to β-actin (n = 5). BMDMs were transfected with Si-scramle (20 nM) and Si-sqstm1 (20 nM) for 48 h, then BMDMs were treated with single LPS or LPS + rhFGF21 for 12 h (n = 4). (F) The expression of p-ULK1 (Ser757) and p-p62 (Ser405) in lysates of BMDMs in indicated groups, and normalized to ULK1 and p62 (n = 4). (G, H) Representative immunofluorescence and quantification of HIF-1α-F4/80 double positive macrophages in liver from mice in indicated groups (n = 5). DAPI (blue), HIF-1α (red), and F4/80 (green). White triangles indicate HIF-1α-F4/80 double positive macrophages. Scale bars: 50 μm. (I) The expression of HIF-1α, LC3 and p62 in Kupffer cells isolated from liver of mice in indicated groups (n = 5). β-actin was used as the standard for verifying equivalent loading. (J) Real-time PCR analysis for hk2, pkm2, and ldha in liver from mice in indicated groups. Data are presented as mean ± SEM of each group. And one-way ANOVA test is using to assess the statistical differences in multiple groups. ns = no significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
2). AFF4 regulates osteogenic potential of human periodontal ligament stem cells via mTOR-ULK1-autophagy axis. Cell proliferation, 2024 (PubMed: 37731335) [IF=8.5]
3). Exosomes derived from miR-26a-modified MSCs promote axonal regeneration via the PTEN/AKT/mTOR pathway following spinal cord injury. Stem Cell Research & Therapy, 2021 (PubMed: 33820561) [IF=7.5]

Application: WB    Species: rat    Sample:

FIGURE S1 | Evaluation of autophagic activity in a SCI rat model treated with exosomes. (a, b) Representative images of western blots used to determine the expression levels of AMPK, p-AMPK, S6K, p-S6K, ULK1, p-ULK1, and p62 and semiquantification of the data. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the control group by t test or ANOVA. #P < 0.05 compared with the Exos group by t test. n = 3 for each group.
4). Taurine alleviates Streptococcus uberis-induced inflammation by activating autophagy in mammary epithelial cells. Frontiers in Immunology, 2021 (PubMed: 33777017) [IF=7.3]

Application: WB    Species: bovine    Sample: MAC-T cells

Figure 2 Taurine treatment decreases mTOR activity and inhibits phosphorylation of ULK1 and ATG13 in MAC-T cells. (A–D) Immunoblots of total protein from MAC-T cells treated with 70 mmol·L−1 taurine for different durations. Quantification of the ratio between the total and phosphorylated (p–) proteins, as determined by densitometric scanning of immunoblots (n = 3). Data are presented as mean ± SEM. (E–G). Cells were incubated with 50 nmol·L−1 Baf-A1 for 1 h, and then treated with 70 mmol·L−1 taurine for 4 h or 100 nmol·L−1 Rapa for 2 h or both; all cells were then probed for the total and phosphorylated (p–) proteins shown. Data are presented as mean ± SEM. *P < 0.05 (significantly different) between the indicated groups; #P < 0.05 (significantly different) between the corresponding negative control groups.
5). α-Hydroxyisocaproic Acid Decreases Protein Synthesis but Attenuates TNFα/IFNγ Co-Exposure-Induced Protein Degradation and Myotube Atrophy via Suppression of iNOS and IL-6 in Murine C2C12 Myotube. Nutrients, 2021 (PubMed: 34371902) [IF=5.9]

Application: WB    Species: Mice    Sample:

Figure 2 The effects of HICA on the intracellular signaling pathways. A typical image for a capillary immunoassay is shown (A). The phosphorylation levels of (B) p70S6K and 4E-BP1; (C) AMPK, ACC, and ULK1; (D) ERK1/2; (E) p38MAPK; and (F) eEF2 are shown. The phosphorylation is normalized to the total protein expression. The β-tubulin content in the lysate was measured as a loading control (G). The time course of these experiments is shown in the upper region. DM: differentiation medium, DMEM: Dulbecco’s modified Eagle’s medium, and w/o AA: without amino acids. Data are displayed as the means ± SD, and n = 4 for each group in all bar graphs. * p < 0.05 and ** p < 0.01 vs. the vehicle-treated group.
6). Icariin exerts anti-tumor activity by inducing autophagy via AMPK/mTOR/ULK1 pathway in triple-negative breast cancer. Cancer cell international, 2024 (PubMed: 38355608) [IF=5.8]
7). Hyperglycemia Aggravates Periodontitis via Autophagy Impairment and ROS-Inflammasome-Mediated Macrophage Pyroptosis. International Journal of Molecular Sciences, 2023 (PubMed: 37047282) [IF=5.6]
8). Radiosensitizing effects of curcumin alone or combined with GLUT1 siRNA on laryngeal carcinoma cells through AMPK pathway-induced autophagy. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, 2021 (PubMed: 33955148) [IF=5.3]
9). Pelargonic acid vanillylamide alleviates hepatic autophagy and ER stress in hepatic steatosis model. Food and Chemical Toxicology, 2023 (PubMed: 37611858) [IF=4.3]
10). Rapamycin Supplementation May Ameliorate Erectile Function in Rats With Streptozotocin-Induced Type 1 Diabetes by Inducing Autophagy and Inhibiting Apoptosis, Endothelial Dysfunction, and Corporal Fibrosis. Journal of Sexual Medicine, 2018 (PubMed: 30224017) [IF=3.5]

Application: WB    Species: rat    Sample: corpus cavernosum

Figure 5.| Autophagy level of the corpus cavernosum (control, n ¼ 6; DMED, n ¼ 6; DMED þ rapamycin, n ¼ 6). A, Representative photomicrographs of LC3B using immunofluorescence (magnification 100). B, Representative Western blot for ULK1, p-ULK1(Ser757),p-ULK1(Ser555), Beclin1, P62, and LC3B (I, II).
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