Rhein (purity: >98% assessed by HPLC) was purchased from Harvey Biotech Co., Ltd. (Beijing, China). For in vitro experiments, rhein was dissolved in dimethyl sulfoxide (DMSO). CNBr Sepharose 4B beads were obtained from GE Healthcare (Piscataway, NJ, USA). The primary antibodies PI3K, p-Akt, Akt, p-mTOR, mTOR, cleaved caspase3, p53, p-p53, Bax, p70 S6 Kinase (p70S6K), p-p70 S6 Kinase (p-p70S6K), cyclin-dependent kinase (CDK) 2, cyclin D1, cyclin E1, E-cadherin, N-cadherin, Heat shock protein 90 (HSP90), 4EBP1, and p-4EBP1 were purchased from Cell Signaling Technology (Beverly, MA, USA). β-Actin, Heat shock factor 1 (HSF1), and cyclin A1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Ki-67 and vimentin were purchased from Abcam (Cambridge, MA, USA).
P70 s6 kinase p70s6k
Manufactured by Cell Signaling Technology
Sourced in United States
P70 S6 Kinase (p70S6K) is a serine/threonine protein kinase that plays a key role in the regulation of cell growth, proliferation, and protein synthesis. It is a downstream effector of the PI3K/Akt signaling pathway and is involved in the phosphorylation of the ribosomal protein S6, which is important for the initiation of protein translation.
Automatically generated - may contain errors
Lab products found in correlation
P akt, by Cell Signaling Technology (2 mentions)
P mtor, by Cell Signaling Technology (2 mentions)
P 4ebp1, by Cell Signaling Technology (2 mentions)
Enhanced chemiluminescence system, by Cytiva (2 mentions)
Horseradish peroxidase conjugated goat anti mouse igg or anti rabbit igg, by Jackson ImmunoResearch (2 mentions)
Hif 1α, by Novus Biologicals (2 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Cyclin a1, by Santa Cruz Biotechnology (1 mentions)
Cnbr sepharose 4b beads, by GE Healthcare (1 mentions)
Cyclin e1, by Cell Signaling Technology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
N cadherin, by Cell Signaling Technology (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Abcam (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
β actin ac 15, by Merck Group (1 mentions)
Phospho akt, by Santa Cruz Biotechnology (1 mentions)
Vascular endothelial growth factor (vegf), by Novus Biologicals (1 mentions)
Phospho mtor, by Cell Signaling Technology (1 mentions)
5 protocols using p70 s6 kinase p70s6k
1
Rhein Modulates PI3K/Akt/mTOR Pathway
2
Protein Expression Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protein content in each sample was determined using a BCA protein assay (Pierce Biotechnology). Proteins were fractionated by SDS-PAGE, transferred onto Hybond enhanced chemiluminescence nitrocellulose membranes (Amersham, Little Chalfont), and probed with antibodies against HIF-1α (Novus Biologicals, Littleton, CO), VEGF (Novus Biologicals), the mammalian target of rapamycin (mTOR) (Cell Signaling, Danvers, MA, USA), phospho-mTOR (Cell Signaling), AKT (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA), phospho-AKT (Santa Cruz Biotechnology, Inc.), p70 S6 kinase (p70S6K) (Cell Signaling), phospho-p70S6K (Cell Signaling), or monoclonal antibodies against β-actin (AC-15, Sigma Aldrich). Horseradish peroxidase-conjugated goat anti-mouse IgG or anti-rabbit IgG (Jackson, West Grove, PA, USA) was used as the secondary antibody, and protein-antibody complexes were visualized by an enhanced chemiluminescence system (Amersham). The signals were quantified with ImageJ software (rsbweb.nih.gov/ij/ ) [39 (link)].
3
Inhibition of PDGF-induced signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Telmisartan, losartan, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Cayman Chemicals (Ann Arbor, MI, USA). Fimasartan was a kind gift from Boryung Pharmaceuticals (Seoul, Korea). GW9662 and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Compound C was purchased from Calbiochem (Darmstadt, Germany). Recombinant rat platelet-derived growth factor (PDGF)-BB protein was purchased from Novus Biologicals (Centennial, CO, USA). Antibodies against mammalian target of rapamycin (mTOR), p-mTOR-Ser2448, p70 S6 kinase (p70S6K), p-p70S6K-Thr389, AMPK, p-AMPK-Thr172, acetyl-CoA carboxylase (ACC), and p-ACC-Ser79 were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody against β-actin was purchased from Sigma-Aldrich. Dulbecco's modified Eagle's medium (DMEM) was obtained from Fisher Scientific (Ottawa, Canada). Dulbecco's phosphate-buffered saline (DPBS), fetal bovine serum (FBS), penicillin and streptomycin antibiotics, trypsin–EDTA solution, and plasticware for cell culture were purchased from Gibco-BRL (Gaithersburg, MD, USA). All other chemicals used were of the purest analytical grade available.
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
The protein sample concentration was measured by Bicinchoninic Acid Protein Assay (ThermoFisher, Waltham, MA, USA). Proteins were subjected to SDS-PAGE separation, and the separated proteins were transferred to PVDF membrane (Pall Corporation, East Hills, NY, USA). The membrane was blocked with primary antibodies p62 (Novus Biologicals, Littleton, CO, USA), LC3 (Novus Biologicals), beclin-1 (Novus Biologicals), p70S6 kinase (p70S6K) (Cell Signaling, Danvers, MA, USA), AKT (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA), mTOR (Cell Signaling), phosphor-AKT (Santa Cruz Biotechnology, Inc.), phosphor-mTOR (Cell Signaling), phosphor-p70S6K (Cell Signaling) or monoclonal antibodies against b-actin (AC-15, Sigma Aldrich). Horseradish peroxidase-conjugated goat anti-mouse IgG or anti-rabbit IgG (Jackson, West Grove, PA, USA) was used as the secondary antibody and protein-antibody complexes were visualized by enhanced chemiluminescence system (Amersham). The signals were quantified with ImageJ software (rsbweb.nih.gov/ij).
5
Western Blot Profiling of Metabolic Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following primary antibodies were used for western blotting: protein kinase B (Akt) (9272; Cell Signaling Technology), p‐Akt (#4060S; Cell Signaling Technology), mechanistic target of rapamycin (mTOR) (#2983; Cell Signaling Technology), p‐mTOR (#2971; Cell Signaling Technology), p70 S6 kinase (p70S6 K) (#9202; Cell Signaling Technology), p‐p70S6 K (#9205; Cell Signaling Technology), 4E‐binding protein 1 (4E‐BP1) (#9452; Cell Signaling Technology), p‐4E‐BP1 (#9459; Cell Signaling Technology), S6 ribosomal protein (S6) (#2217; Cell Signaling Technology), p‐S6 (#4858S; Cell Signaling Technology), glycogen synthase kinase 3β (GSK3β) (#12456; Cell Signaling Technology, Danvers, MA, USA),p‐GSK3β (#5558; Cell Signaling Technology), hypoxia inducible factor 1‐α (HIF‐1α) (NB100‐105; Novus Biologicals), hexokinase (HK) (sc‐374091; Santa Cruz), phosphofructkinase (PFK) (sc‐97025, Santa Cruz), monocarboxylate transporter 1 (MCT1) (QUIAGEN), MCT4 (Quiagen), and oxidative phosphorylation (OXPHOS) (ab110413; Abcam).
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!