Human LAD cell lines SPC-A1 and H1299 were purchased from the Tumor Cell Bank of Chinese Academy of Medical Science (Shanghai, China) and cultured in RPMI 1640 medium containing 10% fetal bovine serum and ampicillin and streptomycin at 37°C in a humidified atmosphere of 95% air and 5% CO2. Docetaxel-resistant SPC-A1 and H1299 cell lines (SPC-A1/DTX and H1299/DTX) were established and preserved in 50 μg/L final concentration of docetaxel in our laboratory. Antibodies against GAPDH, LC3, p62, caspase3, activated (cleaved) caspase3 (c-caspase3), PARP, cleaved PARP (c-PARP), Atg5, HMGB1, mTOR, phosphorylated mTOR (p-mTOR), Akt, p-Akt, S6RP, p-S6RP, MEK, ERK1/2, p-ERK1/2, and H2A were obtained from Cell Signaling Technology. Bafilomycin A1, 3-methyladenine (3-MA), ethyl pyruvate (EP) and PD098059 were purchased from Sigma Aldrich (St. Louis, MO).
Phosphorylated mtor p mtor
Manufactured by Cell Signaling Technology
Sourced in United States
Phosphorylated mTOR (p-mTOR) is a protein that is part of the mammalian target of rapamycin (mTOR) signaling pathway. It plays a key role in the regulation of cellular processes such as cell growth, proliferation, and metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Erk1 2, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
P erk1 2, by Cell Signaling Technology (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
Cleaved parp c parp, by Cell Signaling Technology (1 mentions)
Pd098059, by Merck Group (1 mentions)
Ps6rp, by Cell Signaling Technology (1 mentions)
C caspase 3, by Cell Signaling Technology (1 mentions)
Hmgb1, by Cell Signaling Technology (1 mentions)
3 methyladenine 3 ma, by MedChemExpress (1 mentions)
P70s6k, by Cell Signaling Technology (1 mentions)
Phosphorylated 4ebp1 p 4ebp1, by Cell Signaling Technology (1 mentions)
Ferrostatin 1 fer 1, by MedChemExpress (1 mentions)
Mhy1485, by MedChemExpress (1 mentions)
Srebf1 antibody, by Proteintech (1 mentions)
Phosphorylated akt p akt, by Cell Signaling Technology (1 mentions)
4e bp1, by Cell Signaling Technology (1 mentions)
Z vad fmk, by MedChemExpress (1 mentions)
Rapamycin, by MedChemExpress (1 mentions)
Enhanced chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
4 protocols using phosphorylated mtor p mtor
1
Docetaxel Resistance in Lung Cancer Cell Lines
2
Lipid Metabolism Regulation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RSL3 (#SML2234), aspirin (#A2093), OA (#O1383), POA (#P9417), PA (#P0500) and SA (#S4751) were purchased from Sigma-Aldrich (Darmstadt, Germany). Ferrostatin-1 (Fer-1, #HY100579), Z-VAD-FMK (#HY16658B), 3-Methyladenine (3-MA, #HY19312), MHY1485 (#HY-B0795) and Rapamycin (#HY10219) were obtained from MedChemExpress (NJ, USA). Primary antibodies against AKT (#4691), phosphorylated AKT (p-Akt; #4060), mTOR (#2983), phosphorylated mTOR (p-mTOR; #5536), p70S6K (#2708), phosphorylated p70S6K (p-p70S6K; #9204), 4E-BP1 (#9644), phosphorylated 4E-BP1 (p-4E-BP1; #2855) and SCD1 (#2794) were purchased from Cell Signaling Technology (MA, USA). SREBF1 antibody (#14088-1-AP) was obtained from Proteintech (IL, USA). The antibodies against Ki67 (#ab16667) and 4-HNE (#ab46545) were purchased from Abcam (MA, USA).
3
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RIPA buffer with protease inhibitor cocktail, PMSF (0.1 mM) and NaF (10 mM) was used to lyse cells treated with JQ1 or transfected with siRNA. Protein concentrations were determined by the BCA protein assay kit (Beyotime, Beijing, China). Equal amounts of protein (30 μg) were separated by SDS-PAGE on a 10% gel and transferred to a nitrocellulose membrane. After blocking with 5% defatted milk for 1 h at room temperature, the membrane was incubated overnight at 4 ℃ with primary antibodies against BRD4 (Abcam), c-Myc, CDK4, Bax (Santa Cruz, CA, USA), β-Actin, cleaved-Caspase-3 (cl-Caspase3), PARP, cleaved-PARP (cl-PARP), CDC25A, CyclinD1, P21, Rb, phosphorylated Rb (p-Rb), PI3K, AKT, phosphorylated AKT (p-AKT), mTOR, phosphorylated mTOR (p-mTOR) (Cell Signaling Technology, MA, USA). Secondary antibodies were purchased from Cell Signaling. Protein bands were detected using an enhanced chemiluminescent substrate (Thermo Fisher Scientific Inc.). ImageJ software (National Institutes of Health, USA) was used to quantify the immunoblots. β-Actin served as a control.
4
Western Blot Analysis of Autophagy Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells and liver tissues were harvested and lysed with radioimmunoprecipitation assay buffer, and then the protein concentrations were determined and quantified by bicinchoninic acid assay. Proteins were separated by 12% sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane (Millipore). After blocking with 5% BSA, the membranes were incubated with primary antibodies: anti‐INOS, anti‐LC3A/B, anti‐P62, anti‐mammalian target of rapamycin (mTOR), and phosphorylated‐mTOR (P‐mTOR) (1:1000 dilution; Cell Signaling Technology); anti‐β‐actin (1:1000 dilution; Proteintech); and anti‐GAPDH (1:5000 dilution; OriGene). After washing with Tris‐buffered saline with Tween 20, the membranes were incubated with peroxidase‐conjugated secondary antibodies (goat anti‐rabbit immunoglobulin G [IgG] or goat anti‐mouse IgG; Proteintech). Bound peroxidase activity was detected with an Enhanced Chemiluminescence Detection System (ECL, Thermo Fisher Scientific) using ImmobilonTM Western Chemiluminescent Horseradish Peroxidase Substrate (Merck Millipore).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!