Rabbit anti p s6k t398

Manufactured by Cell Signaling Technology

Rabbit anti-P-S6K T398 is a polyclonal antibody that recognizes the phosphorylated form of the ribosomal protein S6 kinase at threonine 398. This antibody can be used to detect the activation of the S6 kinase protein, which is involved in the regulation of protein synthesis and cell growth.

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4 protocols using rabbit anti p s6k t398

Antibody Immunization and Characterization

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Following Antibodies were used in this study: guinea-pig anti-CG32732 (dSETD3) was self-made in out laboratory by immunization of guinea-pigs with full length 6xHis-CG32732. Rat anti-HA tag (clone 3F10, Roche, 11 867 423 001), rabbit anti-pS6K(T398) (Cell Signaling, 9209), self-made guinea pig anti-S6K [47 (link)], rabbit anti-ERK1/2 (Cell Signaling, 4695).

Tiebe M., Lutz M., Levy D, & Teleman A.A. (2018). Phenotypic characterization of SETD3 knockout Drosophila. PLoS ONE, 13(8), e0201609.

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Western Blot Analysis of Signaling Pathways

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Rabbit anti-BMP2 antibody was obtained from ABclonal Technology. Mouse anti-GAPDH (for normalized control), rabbit anti-mTOR, rabbit anti-P-mTOR, rabbit anti-S6K, rabbit anti-P-S6K (T398), rabbit anti-4E-BP1 and rabbit anti-P-4E-BP1 were from Cell Signaling Technology (CST). Mouse anti-E-cadherin, anti-N-cadherin, anti-β-catenin and anti-vimentin were from BD Biosciences. Both anti-rabbit and anti-mouse labeled with horseradish peroxidase (HRP) were from CST.

Wang M.H., Zhou X.M., Zhang M.Y., Shi L., Xiao R.W., Zeng L.S., Yang X.Z., Zheng X.F., Wang H.Y, & Mai S.J. (2017). BMP2 promotes proliferation and invasion of nasopharyngeal carcinoma cells via mTORC1 pathway. Aging (Albany NY), 9(4), 1326-1339.

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Drosophila Ovary Protein Quantification

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Drosophila ovaries were homogenized in RIPA buffer containing Complete Protease Inhibitors and Phosphatase Inhibitors (Roche). Antibodies were used at the following concentrations: rabbit anti-P-S6K T398 (1:1,000) (Cell Signaling), guinea pig anti-S6K (1:10,000) (PMID: 20444422), guinea pig anti-Nprl3 (1:1,000) (37 (link)), mouse anti-actin (1:10,000)(Abcam), rabbit anti-GFP¹¹ (1:1,000) (Bonopus), rat anti-OLLAS (1:1,000) (Novus), mouse anti-T7 (1:1,000) (Millipore sigma), and rabbit anti-GFP (1:1,000) (Cell signaling). The band intensity was quantified using Image J analysis tool (NIH).

Zhang Y., Ting C.Y., Yang S., Reich J., Fru K, & Lilly M.A. (2022). Wdr59 promotes or inhibits TORC1 activity depending on cellular context. Proceedings of the National Academy of Sciences of the United States of America, 120(1), e2212330120.

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Protein Expression and Quantification

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Whole flies or HeLa cells were lysed in RIPA buffer containing complete protease inhibitors and phosphatase inhibitors (Roche). Western blots were performed as described previously [18 ]. Antibodies were used at the following concentrations: rabbit anti-P-S6K T398 at 1:1000 (Cell Signaling), guinea pig anti-S6K at 1:10,000 (24), mouse anti-actin at 1:10,000 (Abcam), rabbit anti-LC3A/B at 1:1000 (Cell Signaling), rabbit anti-P-S6K T389 at 1:1000 (Cell Signaling), rabbit anti-S6K at 1:1000 (Cell Signaling), rabbit anti-GAPDH at 1:3000 (Cell Signaling), rabbit anti-P-4E-BP1at 1:1000 (Cell Signaling), rabbit anti-4E-BP1 at 1:1000 (Cell Signaling), rabbit anti-GFP at 1:500 (Cell Signaling), rabbit anti-SQSTM1/p62 at 1:500 (Cell Signaling), Mouse anti- SQSTM1/p62 at 1:1000 (Novus Biologicals), rabbit anti-WDR24 at 1:1000 (Novus Biologicals) and goat anti-Cathepsin D at 1:500 (Santa Cruz). The band intensity was quantified using Image J analysis tool (NIH).

Cai W., Wei Y., Jarnik M., Reich J, & Lilly M.A. (2016). The GATOR2 Component Wdr24 Regulates TORC1 Activity and Lysosome Function. PLoS Genetics, 12(5), e1006036.

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