Transfected cells were harvested 48 h after transfection. Cells were lysed in RIPA buffer (Heart, Beijing, China). Total protein samples were separated by SDS‐PAGE polyacrylamide gel and transferred onto a PVDF membrane (Millipore, Billerica, MA, USA). The membranes were immunological overnight at 4°C with primary antibodies as follows: PTEN (1:500; WanleiBio, Shenyang, Liaoning, China); Akt and phosphorylated‐Akt (phosphoresce on S473; 1:500, Abcam, San Francisco, CA, USA); mTOR and phosphorylated‐mTOR (phosphorylated on S2998; 1:1000, Bioworld, Nanjing, Jiangsu, China); Bax and Bcl‐2 (1:1000, Cell Signaling, Danvers, MA, USA); caspase‐9 (1:500, Protein Tech, Wuhan, Hubei, China); and human β‐actin (1:5000; Transgene, China). PVDF membranes were washed with TBST and then incubated with a secondary antibody, HRP‐conjugated goat IgG (1:5000; Transgene, China) for 1 h at 37°C. Signals were detected by the Bio‐Rad Gel imaging system. The images were quantified by Quantity One (Bio‐Rad, USA), and relative protein expression was normalized to β‐actin levels in each sample. All experiments were performed in triplicate.
Mtor and phosphorylated mtor
Manufactured by Bioworld Technology
Sourced in United States, China
MTOR and phosphorylated-mTOR are proteins involved in the regulation of cellular processes. MTOR is a serine/threonine protein kinase that acts as a master regulator of cell growth, proliferation, and metabolism. Phosphorylated-mTOR refers to the active, phosphorylated form of MTOR. These proteins play a key role in various cellular pathways and are important targets for biomedical research.
Automatically generated - may contain errors
Lab products found in correlation
Human β actin, by Transgene (2 mentions)
Akt and phosphorylated akt, by Abcam (1 mentions)
Bax and bcl 2, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Gel imaging system, by Bio-Rad (1 mentions)
Quantity one, by Bio-Rad (1 mentions)
β actin, by Bio-Rad (1 mentions)
Caspase 9, by Proteintech (1 mentions)
Caspase 3, by Abcam (1 mentions)
Flotillin 1, by Abcam (1 mentions)
N cad, by Cell Signaling Technology (1 mentions)
E cad, by Cell Signaling Technology (1 mentions)
Tsg101, by Abcam (1 mentions)
2 protocols using mtor and phosphorylated mtor
1
Western Blot Analysis of Transfected Cells
2
Protein Expression and Signaling Analysis
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Protein extraction and Western blots were conducted and analyzed as previously described [8 (link)] with the antibodies against the following: PTEN, N-Cad, and E-Cad (1:1000; Cell Signaling Technology, MA, USA); Akt and phosphorylated-Akt (phosphorylated on S473), Caspase 3 (1:1000; Abcam, CA, USA); P70S6K and phosphorylated-P70S6K (phosphorylated on S371), mTOR and phosphorylated-mTOR (phosphorylated on S2998; 1:1000; Bioworld, Jiangsu, China); CD63, TSG-101, and Flotillin-1 (1:500; Abcam, CA, USA), and human β-actin (1:5000; Transgene, Beijing, China). The blots were quantified by density relative to β-actin, while the phosphorylation of Akt, mTOR, and P70S6K was determined by density relative to total Akt, mTOR, and P70S6K, respectively. All experiments were performed in triplicate.
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