TEOA was obtained from college of pharmacy, Zhejiang University (Zhejiang, China) and dissolved in DMSO. N-acetylcysteine (NAC), Chloroquine (CQ), 3-Methyladenine (3MA) and Rapamycin were purchased from Medchem Express (MCE, United States). DAPI was purchased from Beyotime (Shanghai, China). The following primary antibodies were used: anti-GAPDH (Abcam ab181602) (1:2,000), anti-Catalase (Abcam ab76024) (1:1,000), anti-MnSOD (Abcam ab68155) (1:2,000), anti-LC3 (Sigma L7543) (1:1,000), anti-p-mTOR (Abcam ab109268) (1:2,000), anti-mTOR (Abcam ab134903) (1:1,000), anti-p-p70S6K (Abcam ab131436) (1:1,000), anti-p70S6K (Abcam ab32529) (1:1,000), anti-p-S6 (CST 4858S) (1:2,000), anti-S6 (CST 2217s) (1:1,000), anti-p-4EBP1 (CST 9459S) (1:1,000), anti-4EBP 1 (CST 9644S) (1:1,000), anti-ATG5 (Abcam ab68155) (1:2,000). The related HRP-conjugated secondary antibody was purchased from Beyotime (Shanghai, China).
Anti p s6
Manufactured by Abcam
Sourced in United States
Anti-p-S6 is an antibody that detects phosphorylated ribosomal protein S6. Ribosomal protein S6 is a component of the 40S subunit of the eukaryotic ribosome and its phosphorylation is associated with the regulation of protein synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Anti s6, by Abcam (3 mentions)
Anti atg5, by Abcam (2 mentions)
3 methyladenine, by MedChemExpress (1 mentions)
N acetylcysteine, by MedChemExpress (1 mentions)
Anti p 4e bp1, by Abcam (1 mentions)
Anti p p70s6k, by Abcam (1 mentions)
Anti p mtor, by Abcam (1 mentions)
Anti p70s6k, by Abcam (1 mentions)
Anti 4e bp1, by Abcam (1 mentions)
Anti mnsod, by Abcam (1 mentions)
Hrp conjugated secondary antibody, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Chloroquine, by MedChemExpress (1 mentions)
Anti lc3, by Merck Group (1 mentions)
Anti mtor, by Abcam (1 mentions)
Anti catalase, by Abcam (1 mentions)
Rapamycin, by MedChemExpress (1 mentions)
Gapdh, by Abcam (1 mentions)
Anti p akt t308, by Abcam (1 mentions)
3 protocols using anti p s6
1
Elucidating Autophagy Regulation by TEOA in Cells
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
After treatment, cells were harvested and lysed by RIPA buffer (Invitrogen). Total protein was measured by BCA protein assay kit (Beyotime Biotechnology, Shanghai, China), separated on 10% SDS-PAGE, and then transferred onto polyvinylidene fluoride (PVDF) and blocked with 5% skimmed milk for 1 h at room temperature. After three times washing with 1×TBST buffer, the PVDF membrane was incubated with the primary antibodies (anti-S6, 1:1000, Abcam; anti-p-S6, 1:1000, Abcam; anti-AKT, 1:500, Abcam; anti-p-AKT (T308), 1:500, Abcam; anti-p-AKT (S473), 1:1000, Abcam; LC3-I/II, 1:500, Abcam; GAPDH, 1:1000, Abcam; Beclin-1, 1:2000, Abcam; anti-ATG5, 1:1000, Abcam) at 4°C for 24 h. Next, the membrane was washed using 1×TBST and followed by incubation with horseradish peroxidase (HRP)-conjugated secondary antibody at room temperate for 1 h. The protein bands were visualized using enhanced chemiluminescence. GAPDH was used as an internal control.
3
Western Blot Analysis of Sestrin Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was performed using standard procedures. The following antibodies were used: anti-Sesn1 (Sanying, Wuhan, China), anti-Sesn2 (Abcam, Cambridge, MA, USA), anti-Sesn3 (Sanying, China), anti-pAMPK (Cell Signaling Technology, Mountain View, CA, USA), anti-AMPK, anti-pS6, anti-S6, anti-LC3 (Abcam, Cambridge, MA, USA), and anti-Gapdh (Xianzhi, Hangzhou, China). Horseradish peroxidaseconjugated secondary antibodies were used and specific antibody-antigen complexes were detected using a chemiluminescent substrate.
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!