Transfected PTC cells were lysed in RIPA buffer (Solarbio, China), and phenylmethylsulfonyl chloride was used as a protease inhibitor to stabilize the whole lysate. The extracted proteins were quantified by the bicinchoninic acid assay (Thermo Scientific, United States). Then the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (BioRad, United States) followed by transferring them to the polyvinylidene difluoride (PVDF) membranes (Millipore, United States). The primary antibodies were as follows: MVP (16478-1-AP, Proteintech), phospho-AKTSer473 (4060S, Cell Signaling Technology), total-AKT (4691S, Cell Signaling Technology), phospho-mTOR (381557, Zen Bioscience), total-mTOR (380411, Zen Bioscience), Phospho-p44/42 (4370T, Cell Signaling Technology), total-p44/42 (4695T, Cell Signaling Technology), phospho-p38 (4511T, Cell Signaling Technology), total-p38 (8690T, Cell Signaling Technology), and -Actin (AP0060, Bioworld Technology). Primary antibodies were used for immunoblotting at 1:1,000 dilution. The membranes were then incubated with a secondary antibody (ab97047 or ab6728, Abcam). Eventually, proteins were detected by the chemiluminescence kit (Thermo Scientific), and images of the protein bands were quantified by ImageJ software (NIH, United States).
Phospho mtor
Manufactured by ZenBio
Sourced in United States
Phospho-mTOR is a laboratory equipment used to detect and quantify the phosphorylated form of the mammalian target of rapamycin (mTOR) protein. mTOR is a key regulator of cell growth, proliferation, and metabolism, and its phosphorylation is an important indicator of its activation status.
Automatically generated - may contain errors
Lab products found in correlation
Bicinchoninic acid assay, by Thermo Fisher Scientific (2 mentions)
Chemiluminescence kit, by Thermo Fisher Scientific (1 mentions)
Ab97047, by Abcam (1 mentions)
Phospho akt ser473 4060, by Cell Signaling Technology (1 mentions)
Total akt 4691 s, by Cell Signaling Technology (1 mentions)
Total p38, by Cell Signaling Technology (1 mentions)
Phospho p44 42, by Cell Signaling Technology (1 mentions)
Total p44 42, by Cell Signaling Technology (1 mentions)
Imagej software, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
Ab6728, by Abcam (1 mentions)
Sodium dodecyl sulfate polyacrylamide gel electrophoresis, by Bio-Rad (1 mentions)
N acetylcysteine nac, by Merck Group (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Sqstm1 p62, by ZenBio (1 mentions)
Phospho pi3k, by Proteintech (1 mentions)
Caspase 9, by ZenBio (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
2 protocols using phospho mtor
1
Western Blot Analysis of Protein Signaling
2
Investigating Apoptosis Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The following antibodies were used: Ki-67 (Cat# Ab16667) from Abcam (Cambridge, Bratian); GAPDH (Cat# 10494-1-AP), phospho-PI3K (Cat# 310163), AKT (Cat# 10176-2-AP), Phospho-AKT (Cat# 66444-1-Ig), Bax (Cat# 50599-2-Ig), Bcl-2 (Cat# 12789-1-AP), and LC3 (Cat# 14600-1-AP) from Proteintech (USA); mTOR (Cat# 380411), phospho-mTOR (Cat# 385033), SQSTM1/P62 (Cat# 380612), PDK-1 (Cat# 220521), and PDH-E1α (Cat# 385512) from ZENBIO (China); caspase 9 (Cat# 9508), cleaved-caspase 9 (Cat# 52873), caspase 3 (Cat# 9662), cleaved-caspase 3 (Cat# 9664), PARP (Cat# 9532), and cleaved-PARP (Cat# 5625) from Cell Signaling Technology (USA); PI3K(Cat# AF6241) from Affinity (USA); phospho-PDHA1(Cat# Ap1022) from Abclonal (China). N-acetyl cysteine (NAC) (Cat# A7250) was purchased from Sigma (USA). As2O3 (ATO, Cat# H20080664) was purchased from Beijing SL Pharmaceutical Company (China).
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