The whole fish (13 dph) and liver tissues (83 dph) stored at − 80 °C were solubilized in RIPA lysis buffer. The protein content was determined using BCA protein assay kit (Yeasen, China). The proteins were separated on 10% SDS-PAGE gel, and then transferred onto PVDF membrane. Anti-phospho ribosomal protein S6 kinase 1 (S6K1) (Thr389), anti-S6K1, anti-phospho ribosomal protein S6 (S6) (Ser235/236), anti-S6 and anti-phospho Grb10 (Ser476) were purchased from Cell Signalling Technology (USA), anti-β-actin antibody from Bioss (China), anti-β-tubulin antibody from Zoman Biotechnology (China). Blots were probed by goat anti-rabbit and goat anti-mouse second antibody with IR-Dye 680 or 800cw labeled (Licor, USA) at room temperature for 1 h. The membranes were then visualized using a LiCor Odyssey scanner (Licor, USA) and quantified with ImageJ 1.44 software (National Institute of Health, MD). The phosphorylation level of S6 and S6K1 were normalized according to the loading of proteins by expressing the data as a ratio of phospho-S6 and phospho-S6K1 over S6 and S6K1, respectively. Besides, the phosphorylation level of Grb10 were normalized according to the loading of proteins by expressing the data as a ratio of phospho-Grb10 over β-actin.
Anti phospho ribosomal protein s6 s6 ser235 236
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-phospho ribosomal protein S6 (S6) (Ser235/236) is a primary antibody that detects the phosphorylation of ribosomal protein S6 at serine residues 235 and 236. Ribosomal protein S6 is a component of the 40S subunit of the eukaryotic ribosome and its phosphorylation is associated with the regulation of cell growth and protein synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey scanner, by LI COR (3 mentions)
Anti β actin antibody, by Bioss Antibodies (3 mentions)
Anti phospho grb10, by Cell Signaling Technology (3 mentions)
Ir dye 680 or 800cw labeled, by LI COR (3 mentions)
Anti s6, by Cell Signaling Technology (3 mentions)
Bca protein assay kit, by Yeasen (3 mentions)
3 protocols using anti phospho ribosomal protein s6 s6 ser235 236
1
Protein Phosphorylation Analysis in Fish Development
2
Phosphorylation Analysis of S6K1, S6, and Grb10
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The whole sh (13 dph) and liver tissues (83 dph) stored at -80 ℃ were solubilized in RIPA lysis buffer. The protein content was determined using BCA protein assay kit (Yeasen, China). The proteins were separated on 10% SDS-PAGE gel, and then transferred onto PVDF membrane. Anti-phospho ribosomal protein S6 kinase 1 (S6K1) (Thr389), anti-S6K1, anti-phospho ribosomal protein S6 (S6) (Ser235/236), anti-S6 and anti-phospho Grb10 (Ser476) were purchased from Cell Signalling Technology (USA), anti-βactin antibody from Bioss (China), anti-β-tubulin antibody from Zoman Biotechnology (China). Blots were probed by goat anti-rabbit and goat anti-mouse second antibody with IR-Dye 680 or 800cw labeled (Licor, USA) at room temperature for 1 h. The membranes were then visualized using a LiCor Odyssey scanner (Licor, USA) and quanti ed with ImageJ 1.44 software (National Institute of Health, MD). The phosphorylation level of S6 and S6K1 were normalized according to the loading of proteins by expressing the data as a ratio of phospho-S6 and phospho-S6K1 over S6 and S6K1, respectively. Besides, the phosphorylation level of Grb10 were normalized according to the loading of proteins by expressing the data as a ratio of phospho-Grb10 over β-actin.
3
Phosphorylation Analysis of S6K1, S6, and Grb10
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The whole sh (13 dph) and liver tissues (83 dph) stored at -80 ℃ were solubilized in RIPA lysis buffer. The protein content was determined using BCA protein assay kit (Yeasen, China). The proteins were separated on 10% SDS-PAGE gel, and then transferred onto PVDF membrane. Anti-phospho ribosomal protein S6 kinase 1 (S6K1) (Thr389), anti-S6K1, anti-phospho ribosomal protein S6 (S6) (Ser235/236), anti-S6 and anti-phospho Grb10 (Ser476) were purchased from Cell Signalling Technology (USA), anti-βactin antibody from Bioss (China), anti-β-tubulin antibody from Zoman Biotechnology (China). Blots were probed by goat anti-rabbit and goat anti-mouse second antibody with IR-Dye 680 or 800cw labeled (Licor, USA) at room temperature for 1 h. The membranes were then visualized using a LiCor Odyssey scanner (Licor, USA) and quanti ed with ImageJ 1.44 software (National Institute of Health, MD). The phosphorylation level of S6 and S6K1 were normalized according to the loading of proteins by expressing the data as a ratio of phospho-S6 and phospho-S6K1 over S6 and S6K1, respectively. Besides, the phosphorylation level of Grb10 were normalized according to the loading of proteins by expressing the data as a ratio of phospho-Grb10 over β-actin.
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