Unless otherwise specified, all chemicals and reagents were purchased from the Sigma Chemical Company (St. Louis, MO, USA). MGF was purchased from Chemicon International (Temecula, CA, USA). Antibodies to IgG, β-actin, AKT, mammalian target of rapamycin (mTOR), ALP, osteocalcin (OCN), CD44, CD34, MGF receptor, phospho-AKT, phospho-mTOR, and phospho-p90RSK1 (Ser380) were purchased from the Millipore Corporation, USA.
Phospho mtor
Manufactured by Merck Group
Sourced in United States
Phospho-mTOR is a laboratory reagent used to detect and quantify the phosphorylated form of the mammalian target of rapamycin (mTOR) protein. mTOR is a key regulator of cellular growth, proliferation, and metabolism. The Phospho-mTOR product enables researchers to monitor the activation state of the mTOR signaling pathway, which is important in various biological processes and disease states.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Merck Group (2 mentions)
Phospho s6, by Cell Signaling Technology (2 mentions)
Phospho akt, by Merck Group (1 mentions)
Phospho p90rsk1 ser380, by Merck Group (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Phospho pi3k, by Cell Signaling Technology (1 mentions)
Vision works ls 6, by Analytik Jena (1 mentions)
β actin, by Analytik Jena (1 mentions)
Nf κb p65, by Abcam (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
β actin, by Novus Biologicals (1 mentions)
Phospho eif2α, by Cell Signaling Technology (1 mentions)
Ero1 lα 3264, by Cell Signaling Technology (1 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
Tunicamycin, by Merck Group (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
Cisplatin, by Merck Group (1 mentions)
5 protocols using phospho mtor
1
Osteoblast Differentiation Molecular Markers
2
Western Blotting Analysis of Cellular Signaling
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Western blotting was performed according to our previous reported [17 (link)]. Antibodies of RAGE, phospho-mTOR (Millipore, Billerica, MA, USA), Bcl-2, Bax, Akt, phospho-Akt, PI3K, phospho-PI3K (Cell signaling technology, Danvers, MA, USA), β-actin, becline-1, ATG5, LC3 (Novus Biologicals, Littleton, Colorado, USA), and NF-κB p65 (Abcam, Cambridge, UK) were used in this study. The protein-relative intensity were quantified by VisionWorks LS 6.3.3 (UVP, Cambridge, UK) and normalized to the β-actin control.
3
Antibodies and Small Molecules for ER Stress
4
Gedunin Cytotoxicity and Signaling Pathway Analysis
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Gedunin (CAS no. 2753‐30‐2, Cat no. 3387/2 with >98.9% purity) procured from R & D Systems (Minneapolis, MN) was dissolved in DMSO and sterile filtered through a 0.22‐μm filter. MTT (3‐(4, 5‐dimethylthiazol‐2yl)‐2,5‐diphenyltetrazolium bromide) and lactate dehydrogenase (LDH) assay kits were sourced from Promega (Madison, WI). Antibodies were sourced from Cell Signaling Technologies (Danvers, MA). Antibodies directed against EGFR, (#4267 1:1000 dilution), Phospho‐EGF Receptor (Tyr1068) Antibody (#2234, dilution 1:1000), Phospho‐Akt (Ser473) (#4060, dilution 1:1000), Phospho‐mTOR (Ser2448) Antibody (#2971, dilution 1:1000), Phospho‐NF‐κB p65 (Ser536) (93H1) Rabbit mAb (#3033 dilution 1:1000), NF‐κB p65 (D14E12) Rabbit Ab (#8242 dilution 1:1000), Phospho‐p70 S6 Kinase (Thr421/Ser424) Antibody (#9204, dilution 1:1000), EGF Receptor (D38B1) Rabbit mAb (#4267 dilution 1:1000), poly ADP‐ribose polymerase (PARP) Antibody (#9542, dilution 1:1000), Cleaved Caspase‐3 (Asp175) Antibody (#9661 dilution 1: 1000), Bcl‐xL (54H6) Rabbit mAb (#2764 dilution 1:1000), Anti‐Bad Antibody (C‐7): sc‐8044 dilution 1:1000), Anti‐α‐Tubulin antibody, Mouse monoclonal (Sigma‐Millipore (St. Louis, MO. T6199, dilution 1:5000) were used in the Western blot analyses.
5
Quantification of Protein Signaling in Adipose Tissues
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Total protein extracts were obtained from BAT, siWAT, and gWAT. After quantification, 15–20 µg was resolved on SDS-PAGE gels (between 6.5 and 12% depending on protein molecular weight) and transferred to PVDF membranes as described in [24 (link),25 (link)]. Membranes were probed with antibodies for uncoupling protein 1 (UCP1) (ab10983, Abcam, Cambridge, UK), for mTOR pathway: mTOR (SAB4501038; Sigma, Kawasaki, Japan), phospho-mTOR (SAB4504476; Sigma), p70S6K (#9202; Cell Signaling, Danvers, MA, USA), phospho-p70S6K (#9205; Cell Signaling), S6 (#2317; Cell Signaling), phospho-S6 (#2211; Cell Signaling); and internal controls: β-actin (A-5316; Sigma), α-tubulin (T-5168; Sigma). Secondary antibodies were purchased by Dako and used at dilution 1:5000 in 3% BSA in TBS-Tween 0.1%. Protein detection was performed using enhanced chemiluminescence reagent ECL (Amersham Biosciences, Little Chalfont, UK). Quantification and analysis of images were carried out by ImageJ software.
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