Phospho eif4ebp1

Manufactured by Cell Signaling Technology

The Phospho-EIF4EBP1 is a laboratory equipment product that detects the phosphorylation state of the EIF4EBP1 protein. EIF4EBP1 is a regulator of protein translation initiation.

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EIF4EBP1 (10 citations)

3 protocols using phospho eif4ebp1

Western Blot Antibody Panel for Cell Signaling

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Primary antibodies against GFP (Takara Bio, 632380, AB_10013427; 1:5000), G-6-PDH (Sigma-Aldrich, A9521, AB_258454; 1:5000), Adh1 (Millipore, 126745, AB_564196; 1:200000), RPS6 (Cell Signaling Technology, 2217, AB_331355; 1:1000), phospho-RPS6 (Ser^235-236) (Cell Signaling Technology, 4856, AB_2181037; 1:1000), RPS6KB (Cell Signaling Technology, 2708, AB_390722; 1:1000), phospho-RPS6KB (Thr³⁸⁹) (Cell Signaling Technology, 9205, AB_330944; 1:1000), EIF4EBP1 (Cell Signaling Technology, 9452, AB_331692; 1:1000), phospho-EIF4EBP1 (Thr^37/46) (Cell Signaling Technology, 2855, AB_560835; 1:1000), phospho-EIF4EBP1 (Ser⁶⁵) (Cell Signaling Technology, 9451, AB_330947; 1:1000), AKT1 (Cell Signaling Technology, 4691, AB_915783; 1:1000), phospho-AKT(Ser⁴⁷³) (Cell Signaling Technology, 4060, AB_2315049; 1:1000), phospho-AKT(Thr³⁰⁸) (Cell Signaling Technology, 13038, AB_2629447; 1:1000), ULK1 (Cell Signaling Technology, 8359, AB_11178668; 1:1000), phospho-ULK1(Ser⁷⁵⁷) (Cell Signaling Technology, 6888, AB_10829226; 1:1000), MAP1LC3 AB (Cell Signaling Technology, 12741, AB_2617131; 1:1000), β-actin (Cell Signaling Technology, 4967, AB_330288; 1:1000) were used.

Nicastro R., Gaillard H., Zarzuela L., Péli-Gulli M.P., Fernández-García E., Tomé M., García-Rodríguez N., Durán R.V., De Virgilio C, & Wellinger R.E. (2022). Manganese is a physiologically relevant TORC1 activator in yeast and mammals. eLife, 11, e80497.

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Antibodies and Reagents for mTOR Signaling

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Antibodies against HMGA1 (ab129153), HMGA1-ChIP Grade (ab252930), and FKBP12 (ab2918) were purchased from Abcam. Antibodies against HMGA1 (sc-393213) and FKBP12 (sc-133067) and mouse IgG (sc-2025) were purchased from Santa Cruz Biotechnology. Antibodies against S6 ribosomal protein (#64108) and phospho-S6 ribosomal protein (#81736) and rabbit IgG (#2729), mTOR (#2983), phospho-mTOR (#5536), eIF4EBP1(#9452), and phospho-eIF4EBP1(#9456) were purchased from Cell Signaling Technology (Danvers, MA). Antibody against beta actin (81115-1-RR) was purchased from Protein Tech (Wuhan, China). Rapamycin was obtained from MedChemExpress (HY-10219).

Guo J.R., He K.Y., Yuan J.L., An W., Yin W.T., Li Q.T., Lu L.Y., Yang J.Y., Liu M.J., Li Y.J., Zhao Y., Yang Q., Lei X.Y., Gao F., Zhang L., Wu D.H., Li J.Q., Zhao Z.L., Liu H., Zhu L.J., Xiang X.Y., Sun Q.H., Jian Y.P, & Xu Z.X. (2024). HMGA1 sensitizes esophageal squamous cell carcinoma to mTOR inhibitors through the ETS1-FKBP12 axis. International Journal of Biological Sciences, 20(7), 2640-2657.

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Western Blot Analysis of Zebrafish Ganglioneuroma Cells

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Protein was extracted from zebrafish primary ganglioneuroma cells using RIPA buffer after compound treatment for 18 h. Western blotting was performed as described previously (Tao et al., 2013 (link)). Protein samples were separated by a 10% polyacrylamide gel and transferred to a polyvinylidene fluoride membrane (#PI88518; Thermo Fisher Scientific). The membrane was blocked in PBST containing 5% nonfat milk for 1 h at room temperature, incubated with a primary antibody for 2 h diluted in blocking buffer. After washing three times in PBST, the membrane was incubated with a secondary antibody for 1 h diluted in blocking buffer and washed three times again in PBST. The results were visualized on autoradiography films with SuperSignal West Pico Chemiluminescent Substrate (#PI34080; Pierce) or SuperSignal West Dura Extended Duration Substrate (#34075; Pierce). Antibodies against phospho-S6 ribosomal protein (Ser235/236, #2211; Cell Signaling Technology), S6 ribosomal protein (#2317; Cell Signaling Technology), phospho-EIF4EBP1 (Thr37/46, #2855; Cell Signaling Technology), EIF4EBP1 (#9644; Cell Signaling Technology), and α-Tubulin (#T6074; Sigma) were used as primary antibodies.

Tao T., Shi H., Wang M., Perez-Atayde A.R., London W.B., Gutierrez A., Lemos B., Durbin A.D, & Look A.T. (2020). Ganglioneuromas are driven by activated AKT and can be therapeutically targeted with mTOR inhibitors. The Journal of Experimental Medicine, 217(10), e20191871.

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