P ulk1 ser317

Manufactured by Cell Signaling Technology

P-ULK1 (Ser317) is a laboratory product offered by Cell Signaling Technology. It is an antibody that detects phosphorylation of ULK1 at serine 317. ULK1 is a protein kinase that plays a key role in the initiation of autophagy. The phosphorylation of ULK1 at serine 317 is a regulatory event, but the specific functional implications are not detailed here.

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P-ULK1 (48 citations)

2 protocols using p ulk1 ser317

Autophagy and Apoptosis Signaling Pathway

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The whole proteins in each cell samples were extracted by RIPA Lysis Buffer containing 1 mM PMSF (Beyotime). The protein was blocked and incubated with the primary antibodies against beclin-1, p62/SQSTM1 and LC3 (MBL; 1:1000), caspase 3, GAPDH, P-Akt (Ser473), P-AMPK (Thr172), P-ULK1 (Ser317) and P-mTOR (Ser2448) (Cell Signaling; 1:1000), overnight at 4 °C, respectively. The protein was then incubated with HRP-conjugated secondary antibody (1:5000) for 1 h at room temperature. The protein was detected using the eECL Western Blot Kit (Beyotime).

Zhang P., Lai Z.L., Chen H.F., Zhang M., Wang A., Jia T., Sun W.Q., Zhu X.M., Chen X.F., Zhao Z, & Zhang J. (2017). Curcumin synergizes with 5-fluorouracil by impairing AMPK/ULK1-dependent autophagy, AKT activity and enhancing apoptosis in colon cancer cells with tumor growth inhibition in xenograft mice. Journal of Experimental & Clinical Cancer Research : CR, 36, 190.

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Western Blot Analysis of Autophagy Markers

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Mouse monoclonal Abs against LC3, β-Actin, Akt1, and rabbit polyclonal Abs against AnxA2, mTOR, ULK1, p-Akt1, and p-mTOR were obtained from Santa Cruz Biotechnology. Rabbit polyclonal Abs against Fam13A was obtained from Proteintech Group (Chicago, IL), while p-ULK1 (ser317) was obtained from Cell Signaling Technology (Danvers, MA). The samples derived from cells and lung homogenates were lysed in RIPA buffer, separated by electrophoresis on 12% SDS-PAGE gels and transferred to nitrocellulose transfer membranes (GE Amersham Biosciences, Pittsburgh, PA). Proteins were detected by western blotting using primary Abs at a concentration of 1/200 (Santa Cruz Biotechnology) or 1/1000 (Cell Signaling Technology) and were incubated overnight. Labelling of the first Abs was detected using relevant secondary Abs conjugated to HRP (Santa Cruz Biotechnology), detected using ECL regents (Santa Cruz Biotechnology) (38 ). Gel bands were quantified by Quantity One software (Bio Rad) and error bar represents for three independent immunoblotting assays (38 ). Phosphorylated and total protein levels were determined and quantified by three independent successive immunoblotting membranes.

Li R., Tan S., Yu M., Jundt M.C., Zhang S, & Wu M. (2015). Annexin A2 regulates autophagy in Pseudomonas aeruginosa infection through Akt1-mTOR-ULK1/2 signaling pathway. Journal of immunology (Baltimore, Md. : 1950), 195(8), 3901-3911.

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