Total RNA was extracted from mouse skeletal muscle with the use of an RNeasy Fibrous Tissue Mini Kit (Qiagen, Hilden, Germany). Quantitative reverse transcription (RT) and PCR analysis was performed as described previously8 (link), with 36B4 mRNA as an internal control and with the use of an ABI StepOne Plus Real-Time PCR system (Applied Biosystems, Waltham, MA). The sequences of primer pairs are available on request. For immunoblot analysis, the protein concentration of cell or tissue extracts was measured with a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA), and the same amount of protein was loaded in each lane. Primary antibodies included those to the following proteins: PDK1 (#3062), Akt (#9272), Thr308-phosphorylated Akt (#9275), Ser473-phosphorylated Akt (#9271), S6K (#9202), Thr389-phosphorylated S6K (#9205), S6 (#2217), Ser235- and Ser236- phosphorylated S6 (#2211), all from Cell Signaling Technology (Danvers, MA), as well as Thr229-phosphorylated S6K (1015B) from R&D Systems (Minneapolis, MN). Akt activity was measured with an Akt Kinase Assay Kit (#9840, Cell Signaling Technology).
Thr389 phosphorylated s6k
Manufactured by Cell Signaling Technology
Sourced in France
Thr389-phosphorylated S6K is a laboratory product that detects the phosphorylation of the Thr389 residue in the S6 kinase (S6K) protein. S6K is a downstream target of the mTOR signaling pathway, and its phosphorylation at Thr389 is a key indicator of mTOR activity. This product can be used to monitor mTOR signaling in cell-based assays or tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Phosphorylated akt ser473, by Cell Signaling Technology (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Akt kinase assay kit, by Cell Signaling Technology (1 mentions)
Phosphorylated akt thr308, by Cell Signaling Technology (1 mentions)
Abi steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions)
Rneasy fibrous tissue mini kit, by Qiagen (1 mentions)
Rotor gene sybr green pcr master mix, by Qiagen (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Agilent Technologies (1 mentions)
Tri reagent, by Euromedex (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
2 protocols using thr389 phosphorylated s6k
1
Quantitative RT-PCR and Immunoblot Analysis of Mouse Skeletal Muscle
2
Molecular Mechanisms of P38 Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protease inhibitor cocktail, primers, and the antibody against P38 were purchased from Sigma‐Aldrich (Saint‐Quentin‐Fallavier, France); antibodies against 4E‐BP1, Akt, Ser473‐phosphorylated Akt, S6K, and Thr389‐phosphorylated S6K were from Cell Signaling Technology; horseradish peroxidase‐conjugated secondary antibodies were from DAKO (Trappes, France); SuperScript® III reverse transcriptase, random hexamer, and oligo‐dT primers were from Invitrogen (Life Technologies, Saint‐Aubin, France); Tri reagent was from Euromedex (Mundolsheim, France); and Rotor‐Gene SYBR Green PCR Master Mix from Qiagen (Courtaboeuf, France).
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!