NuPAGE Novex Midi Gels (4–12%) from Life Technologies were used for protein separation. The following antibodies were used: Anti-PSD95 antibody (Abcam, Cat. #ab18258), Synaptophysin (D35E4) XP® Rabbit mAb (Cell Signaling, Cat. #5461), Complex II Subunit Monoclonal Antibody (clone 21A11AE7, Life Technology, #439454), Phospho-p70 S6 Kinase (Thr421/Ser424) (Cell Signaling, #9204), S6K(p70) (Cell Signaling, #2217), Phospho-Akt (Ser473) Antibody (Cell Signaling, Cat. #9271s), Akt (pan) (11E7) Rabbit mAb (Cell Signaling, Cat. #4685s), Ube3a (Abgent, #346954), β-Tubulin Antibody (TUB 2.1, Santa Cruz, Cat. #sc-58886) and β-Actin antibody (Sigma, Cat. #A1978). After incubating with corresponding infrared fluorescent secondary antibodies from Li_COR Biosciences (#926–32213 IRDye® 800CW Donkey anti-Rabbit IgG, and #926–68072 IRDye® 680RD Donkey anti-Mouse IgG), the signal of specific protein was detected and quantified on the Odyssey CLx imager (LI-COR).
Phospho p70 s6 kinase thr421 ser424
Manufactured by Cell Signaling Technology
Phospho-p70 S6 Kinase (Thr421/Ser424) is a primary antibody that detects the phosphorylation of p70 S6 Kinase at Thr421 and Ser424 residues. p70 S6 Kinase is a serine/threonine protein kinase that plays a key role in the regulation of cell growth, proliferation, and protein synthesis.
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Lab products found in correlation
Irdye 680rd donkey anti mouse igg, by LI COR (1 mentions)
Phospho akt ser473 antibody, by Cell Signaling Technology (1 mentions)
Akt pan 11e7 rabbit mab, by Cell Signaling Technology (1 mentions)
Ube3a, by Abcepta (1 mentions)
Anti psd 95 antibody, by Abcam (1 mentions)
Odyssey clx imager, by LI COR (1 mentions)
β actin antibody, by Merck Group (1 mentions)
Phospho p70 s6 kinase thr389, by Cell Signaling Technology (1 mentions)
P 4e bp1 t37 46, by Cell Signaling Technology (1 mentions)
Total akt, by Merck Group (1 mentions)
P70 s6 kinase 49d7, by Cell Signaling Technology (1 mentions)
P akt ser473, by Merck Group (1 mentions)
Total 4e bp1, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
2 protocols using phospho p70 s6 kinase thr421 ser424
1
Protein Expression Profiling Assay
2
Protein Signaling Pathway Analysis
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Western blot analysis used specific antibodies for β-actin (Sigma Aldrich), p-Akt (Ser473) (#4060), total Akt (#9272), p-ERK1/ERK2 (Thr202/Tyr204) (#4370), total ERK1/ERK2 (#9102), Phospho-p70 S6 Kinase (Thr421/Ser424) (#9204), Phospho-p70 S6 Kinase (Thr389) (#9205), p70 S6 Kinase (49D7)(#2708), p-4EBP1(T37/46) (#2855) and total 4EBP1 (#9644) (Cell Signaling). MCF-7 cells were seeded at 500.000 cells in 10 cm dishes in growth media. Next day, cells were treated with Veh, 100 nM OA, PA, LA or SA for indicated times. Cell lysate was prepared using RIPA buffer. Samples were sonicated three times 10 seconds to shear the DNA. 10 µg protein was loaded onto 10% SDS gels. Antibodies were used at 1:500 except for β-actin (1:5000). Proteins were visualized using Odyssey LICOR imaging system.
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