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Anti phospho ulk1ser555

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Anti-Phospho-ULK1Ser555 is a primary antibody that recognizes ULK1 (unc-51 like autophagy activating kinase 1) when phosphorylated at serine 555. This antibody can be used to detect the phosphorylation state of ULK1 in biological samples.

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Lab products found in correlation

Anti phospho ampkα thr172, by Cell Signaling Technology (2 mentions) Anti ulk1, by Cell Signaling Technology (1 mentions) Anti gclc, by Abcam (1 mentions) Anti gapdh, by Abcam (1 mentions) Anti mfn2, by Cell Signaling Technology (1 mentions) Anti pgc1 alpha, by Abcam (1 mentions) Anti ampkα, by Abcam (1 mentions) Anti prdx3, by Abcam (1 mentions) Anti citrate synthase, by Abcam (1 mentions) Anti lc3b, by Cell Signaling Technology (1 mentions) Anti p62, by Abcam (1 mentions) Anti nox2 gp91phox, by Abcam (1 mentions) Anti mouse and anti rabbit secondary antibodies, by Bio-Rad (1 mentions) Anti phospho ampk thr172, by Cell Signaling Technology (1 mentions) Anti lc3, by Nanotools (1 mentions) Anti phospho ulk1, by Cell Signaling Technology (1 mentions) Anti sv40 lt, by Santa Cruz Biotechnology (1 mentions) Anti sqstm1 p0067, by Merck Group (1 mentions) Anti ampk, by Cell Signaling Technology (1 mentions) Anti p gp, by Abcam (1 mentions)

3 protocols using anti phospho ulk1ser555

1

Comprehensive Protein Analysis Protocol

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Primary antibodies used for Western blotting are listed as follows, the dilutions are indicated in the list. Anti-PGC1 alpha (ab191838, dilution 1:1000), Anti-NOX2/gp91phox (ab129068, 1:500), Anti-GCLC (ab190685, 1/500), Anti-PRDX3 (ab73349, 1:1000), Anti-p62 (ab91526, 1:1000), Anti-Citrate synthase (ab129095, 1:1000), Anti-GAPDH (ab8245, 1:2000) from Abcam (Cambridge, UK); Anti-AMPKα (#2532, 1:400), Anti- Phospho-AMPKαThr172 (#2535, 1:400), Anti-MFN2 (#9482, 1:500), Anti-ULK1 (#8054, 1:200), Anti-Phospho-ULK1Ser555 (#5869, 1:250), Anti-LC3B (#43566, 1:250), from Cell Signalling (Cell Signalling Technology, Danvers, MA, USA); Anti-SOD2 (ADI-SOD-111, 1:2000) from Enzo Life Sciences (Farmingdale, New York, USA). Primary antibodies Anti-MYH I (BA-D5, 1:200), Anti-MYH IIb (BF–F3, 1:250), and Anti-MYH IIa (SC-71, 1:250) from Developmental Studies Hybridoma Bank (Iowa, USA) and Wheat Germ Agglutinin (WGA), CF®405S Conjugate (29,027–1, 1:200, Generon, Slough, UK) for membrane staining were used for immunohistochemistry.
Ersoy U., Kanakis I., Alameddine M., Pedraza-Vazquez G., Ozanne S.E., Peffers M.J., Jackson M.J., Goljanek-Whysall K, & Vasilaki A. (2023). Lifelong dietary protein restriction accelerates skeletal muscle loss and reduces muscle fibre size by impairing proteostasis and mitochondrial homeostasis. Redox Biology, 69, 102980.
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2

Plasmid Transfection and Antibody Analysis

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The following plasmids were used for transfection: pcDNA3.1(-)-GFP-LC3 [47 (link)], mRFP-GFP-LC3 construct [48 (link)]. hPC1-expressing plasmid WT (REP10) was a gift from Gregory Germino (Addgene plasmid #21368) [49 (link)] as was hPC2-expressing plasmid (M-PKD2 (OF2-3); Addgene plasmid #21370) [21 (link)].
The following siRNAs were used: siRNA against mouse Pkd2 (AM16708, assay ID 150154, Thermo Fischer Scientific), siRNA against mouse BECN1 (4390771, Ambion, Woolston, UK) and negative control DsiRNA (51-01-14-04, IDT).
The following antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA): anti-β-Actin (4970), anti-FoxO1 (14952), anti-ULK1 (8054), anti-phospho-ULK1 (Ser555) (5869), anti-phospho-ULK1 (Ser757) (14202), anti-phospho-ULK1 (Ser317) (3776), anti-AMPK (5832), anti-phospho AMPK (Thr172) (50081), anti-S6 (2217) and anti-p-S6 (4858). Anti-PC1 (sc-130554), anti-PC2 (sc-28331) and anti-SV40 LT were from Santa Cruz Biotechnology (Dallas, TX, USA), anti-Sqstm1 (P0067) and anti-EpCam (HPA026761) from Sigma-Aldrich (Saint-Louis, MO, USA, anti-LC3 from Nanotools (clone 5F10; München, Germany), anti-PgP from Abcam (ab170904; Cambridge, UK) and anti-AQP1 from Novus Biologicals (84488; Abingdon, UK). Secondary anti-rabbit and anti-mouse antibodies were from Bio-Rad Laboratories (Hercules, CA, USA).
Decuypere J.P., Van Giel D., Janssens P., Dong K., Somlo S., Cai Y., Mekahli D, & Vennekens R. (2021). Interdependent Regulation of Polycystin Expression Influences Starvation-Induced Autophagy and Cell Death. International Journal of Molecular Sciences, 22(24), 13511.
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3

AMPK Activation in HEK293 Cells

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HEK293 cells were grown in DMEM media until they reached confluency. Then, cells were washed with KHR/Glu media (125 mM NaCl, 3 mM KCl, 1.5 mM CaCl2, 0.5 mM MgSO4, 0.5 mM KH2PO4, 2.5 mM NaHCO3, 10 mM HEPES, pH: 7.4/ 25 mM Glucose) and maintained in this media supplemented or not with the appropriated concentration of different compounds for 1 h. Phenformin 5 mM and different doses of A-769662 (25 to 100 μM) were used as a positive control of AMPK activation. Then, cell lysis and protein extracts were obtained and analyzed as in Garcia-Haro et al., 201033 (link). In brief, cell extracts (30 μg) were boiled in electrophoresis sample buffer and analyzed by SDS/PAGE and immunoblotting using appropriate antibodies: anti-phospho-AMPKα-Thr172 (#2535), anti-AMPKβ1 (#4182), anti-phospho-ACC-Ser79 (#3661), anti-ACC (#3662), anti-phospho-Raptor-Ser792 (#2083), anti-phospho-ULK1-Ser555 (#5869), anti-phospho-p38-Thr180/Tyr182 (#9211), and anti-phospho-JNK-Thr183/Tyr185 (#4668), were from Cell Signaling Technology (Hertfordshire, UK). Anti-αTubulin (T6199) was from Sigma. Secondary antibodies were from Santa Cruz Biotechnology (Santa Cruz, USA). Immunoblots were analyzed with the ECL+ reagent (GE Healthcare, Barcelona, Spain) and chemiluminescence was detected using a FUJIFILM LAS-3000 lite imager. Quantification of the bands was performed using the Image Studio lite v4.0 software.
Vela M., García-Gimeno M.A., Sanchis A., Bono-Yagüe J., Cumella J., Lagartera L., Pérez C., Priego E.M., Campos A., Sanz P., Vázquez-Manrique R.P, & Castro A. (2021). Neuroprotective effect of IND1316, an indole-based AMPK activator, in animal models of Huntington disease. ACS chemical neuroscience, 13(2), 275-287.
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