Anti s6 54d2

Manufactured by Cell Signaling Technology

Sourced in United States

The Anti-S6 (54D2) is a monoclonal antibody that recognizes the S6 ribosomal protein. This antibody is intended for research use only.

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Lab products found in correlation

Anti ha, by New England Biolabs (1 mentions) Anti deptor d9f5, by Cell Signaling Technology (1 mentions) Anti phospho s6 s240 244, by Cell Signaling Technology (1 mentions) Antiphospho akts473 d9e, by Cell Signaling Technology (1 mentions) Anti gfp 2956, by Cell Signaling Technology (1 mentions) Anti ubiquitin p4d1 sc 8017, by Santa Cruz Biotechnology (1 mentions) Anti akt pan c67e7, by Cell Signaling Technology (1 mentions) Polyvinylidene fluoride membrane, by Bio-Rad (1 mentions) Anti gapdh d16h11, by Cell Signaling Technology (1 mentions) Anti mtor 7c10, by Cell Signaling Technology (1 mentions) Anti coxiv ab16056, by Abcam (1 mentions) Anti gapdh mab374, by Merck Group (1 mentions) Enhanced chemiluminescence reagent kit, by Merck Group (1 mentions) Polyvinylidene fluoride membrane, by Merck Group (1 mentions) R0020, by Solarbio (1 mentions)

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Anti-S6 (87 citations)

3 protocols using anti s6 54d2

Western Blot Analysis of Protein Targets

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Cells were lysed in Laemmli buffer, proteins were separated on SDS-PAGE, and transferred to polyvinylidene fluoride membranes (Bio-Rad). Primary antibodies used were anti-FLAG (2368; NEB; 1:1000 dilution), anti-HA (3724; NEB; 1:1000 dilution), anti-mTOR (7C10; Cell Signaling; 1:1000 dilution), anti-DEPTOR (D9F5; Cell Signaling; 1:1000 dilution), anti-S6 (54D2; Cell Signaling; 1:1000 dilution), anti-phospho-S6^S240/244 (2215; Cell Signaling; 1:1000 dilution), anti-pan-AKT (C67E7; Cell Signaling; 1:1000), antiphospho-AKT^S473 (D9E; Cell Signaling; 1:1000 dilution), anti-ACTIN (D6A8; Cell Signaling; 1:2000 dilution) and anti-GAPDH (D16H11; Cell Signaling; 1:2000 dilution), anti-4G10 (05-321; Millipore–Sigma; 1:1000 dilution), anti-GFP (2956; Cell Signaling; 1:1000 dilution), antiubiquitin P4D1 (sc-8017; Santa Cruz; 1:1000 dilution), anti-EPHB2 D2X2I (83029; Cell Signaling; 1:1000 dilution), anti-EPHB2 (AF467; R&D Systems; 1:200 dilution), and anti-SYK D3ZE1 (13198; Cell Signaling; 1:500 dilution).

M. Gagné L., Morin N., Lavoie N., Bisson N., Lambert J.P., Mallette F.A, & Huot M.É. (2021). Tyrosine phosphorylation of DEPTOR functions as a molecular switch to activate mTOR signaling. The Journal of Biological Chemistry, 297(5), 101291.

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Immunoblotting of Key Erythroid Proteins

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Immunoblotting was performed as previously described (99 (link)). Anti-LAT3 (ab55552), anti-PDHD (ab67592), and anti-COX-IV (ab16056) rabbit polyclonal antibodies were purchased from Abcam (Cambridge, England). Anti-GAPDH (MAB374) and anti-TFRC (136800) mouse monoclonal antibodies were purchased from Millipore and Invitrogen, respectively. Mouse monoclonal anti-ATP6V1H (G-2), anti-TUBA1A (DM1A), and anti-ACTB (C-2) and goat polyclonal anti-FECH (C-20), anti-HSPD1 (K-19), and anti-GATA1 (N-1) antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Rabbit monoclonal anti-LC3B (2775S), anti-(pS51)eIF2α (D9G8), anti-eIF2α (D7D3), anti-(pT37/pT46)4E-BP1 (236B4), anti-(pT389)S6K1 (108D2), anti-4E-BP1 (53H11), anti-4E-BP2 (#2845), anti-S6K1 (49D7), and anti-(pS240/244)S6 (D68F8) and mouse monoclonal anti-S6 (54D2) were purchased from Cell Signaling Technologies (Danvers, MA, USA). The rabbit polyclonal anti-PPOX antibody was a generous gift of Dr. Harry A. Dailey (University of Georgia, GA, USA). The rabbit polyclonal anti-Ankyrin1 and anti-EPB42 antibodies were generous gifts of Dr. Samuel E. Lux IV (Harvard Medical School, MA, USA). The anti-ALAS2 rabbit polyclonal antibody was a generous gift of Dr. Hiroshi Munakata (Kinki University, Osaka-Sayama, Japan).

Chung J., Bauer D.E., Ghamari A., Nizzi C.P., Deck K.M., Kingsley P.D., Yien Y.Y., Huston N.C., Chen C., Schultz I.J., Dalton A.J., Wittig J.G., Palis J., Orkin S.H., Lodish H.F., Eisenstein R.S., Cantor A.B, & Paw B.H. (2015). The mTORC1/4E-BP pathway coordinates hemoglobin production with L-leucine availability. Science signaling, 8(372), ra34.

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Leucine-Induced Signaling Pathway Analysis

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A549 and H1299 cells were seeded in 6-well plates. When the cells reached 80% confluence, they were exposed to leucine (7.5 mM) for 15 min, 30 min, 1 h, 2 h, and 4 h, respectively. Stable A549 and H1299 cell lines were analyzed. The cells were lysed in radioimmunoprecipitation assay buffer (R0020, Solarbio) supplemented with a protease inhibitor cocktail and phosphatase inhibitor mixture. The total protein concentration was measured using a bicinchoninic acid protein assay kit. The proteins were heated at 97°C for 5 min, separated by 10–15% SDS-PAGE, and transferred onto polyvinylidene fluoride membranes (Millipore, USA). The membranes were then incubated with specific primary antibodies and corresponding secondary antibodies. Protein bands were visualized using an enhanced chemiluminescence reagent kit (Millipore). The following antibodies were used: anti-BCKDK (E-12) (sc-374425; Santa Cruz Biotechnology), anti-phospho-S6 (Ser235/236) (D57.2.2E; Cell Signaling Technology), anti-S6 (54D2; Cell Signaling Technology), and anti-Rab1A (G-10) (sc-377201; Santa Cruz Biotechnology). The mean optical densities of the bands were quantified using ImageJ acquisition and analysis software.

Xue M., Xiao J., Jiang W., Wang Y., Zuo D., An H, & Ren L. (2023). Loss of BCAA catabolism enhances Rab1A-mTORC1 signaling activity and promotes tumor proliferation in NSCLC. Translational Oncology, 34, 101696.

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