Rps6 5g10

Manufactured by Cell Signaling Technology

Sourced in United States

RPS6(5G10) is a monoclonal antibody that recognizes the ribosomal protein S6 (RPS6). RPS6 is a component of the 40S ribosomal subunit and is involved in the regulation of cell growth and protein synthesis.

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Lab products found in correlation

3 protocols using rps6 5g10

Immunoblotting of Innate Immune Signaling

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Cells were lysed in RIPA buffer (+ 1x HALT protease and phosphatase
inhibitor and 10–30 μg total protein from whole cell lysates was
run on SDS-PAGE and transferred to PVDF membranes (Thermo Scientific). The
membranes were probed in 5 % BSA or milk in PBS-T (Phosphate-buffered
saline/Tween 20) for ZAP (N3C2, GeneTex), ISG15 (Cell Signaling, #2743), OAS1
(D1W3A, Cell Signaling), RIG-I (Alme-1, AdipoGen), IRF1 (D5E4, Cell Signaling),
IRF3 (D83B9, Cell Signaling), phospho-IRF3 (Ser386) (EPR2346, Abcam), STAT1
(42H3, Cell Signaling), phospho-STAT1 (Tyr701) (58D6, Cell Signaling), RPS6
(5G10, Cell Signaling), CSTF2 (Bethyl Laboratories), Calnexin (C5C9, Cell
Signaling), FLAG (M2, Sigma), Myc (71D10, Cell Siganling) or β-Actin-HRP
(13E5, Cell Signaling). For detailed information about the source of the
antibodies and dilutions used please refer to the Life Sciences Reporting
Summary.

Schwerk J., Soveg F.W., Ryan A.P., Thomas K.R., Hatfield L.D., Ozarkar S., Forero A., Kell A.M., Roby J.A., So L., Hyde J.L., Gale M J.r., Daugherty M.D, & Savan R. (2019). RNA-binding protein isoforms ZAP-S and ZAP-L have distinct antiviral and immune resolution functions. Nature immunology, 20(12), 1610-1620.

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Immunoblot analysis of mTOR signaling

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Cells were washed and scraped with ice-cold PBS. Lysates were prepared using lysis buffer P containing protease and phosphatase inhibitors (#1861280; Thermo Scientific, Dreieich, Germany). 10 μg of protein per condition were subjected to SDS-PAGE analysis. Membranes were probed with antibodies against phospho-4EBP1 (Thr37/46) (236B4; Cell Signaling), phospho-RPS6 (Ser 240/244) (D68F8; Cell Signaling), phospho-RPS6 (Ser 235/236) (D57.2.2.E; Cell Signaling), RPS6 (5G10; Cell Signaling), 4EBP1 (53H11; Cell Signaling) or actin (# sc-1616 Santa Cruz Biotechnology, Dallas, Texas, USA). The secondary anti-rabbit and anti-goat antibodies were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Chemiluminescence solution was used for detection and was set up with 1 ml solution A (200 ml 0.1 M Tris-HCl pH 8.6, 50 mg Luminol), 100 μl solution B (11 mg p-hydroxycurmarinacid, 10 ml DMSO) and 0.3 μl H₂O₂ (30%).

Harter P.N., Jennewein L., Baumgarten P., Ilina E., Burger M.C., Thiepold A.L., Tichy J., Zörnig M., Senft C., Steinbach J.P., Mittelbronn M, & Ronellenfitsch M.W. (2015). Immunohistochemical Assessment of Phosphorylated mTORC1-Pathway Proteins in Human Brain Tumors. PLoS ONE, 10(5), e0127123.

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Immunoblotting Analysis of CIT Compound Effects

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Antibodies to detect STMN1 (D1Y5A, #13655), phospho-STMN1 (Ser16, #3353), CRMP2 (D8L6V, #35672), phospho-CRMP2 (Thr514, #9397), PARP (46D11, #9532), c-Jun (60A8, #9165), phospho-c-Jun (Ser63, #91952), AKT (C67E7, #4691), phospho-AKT (Ser473, D9E, #4060), ERK1/2 (137F5, #4695), phospho-ERK1/2 (Thr202/Tyr204, #4370), RPS6 (5G10, #2217), phospho-RPS6 (Ser235/236, #4858) were from Cell Signaling (Danvers, MA, USA). Anti-β-Actin (AC-15, A5441) and Anti-α-Tubulin (DM1a, MABT205) were from Sigma-Aldrich.
CIT compounds CIT-026 and CIT-223 were synthesized by the aldol condensation between a tetralone and an indole aldehyde as described previously (19 (link)). CITs were dissolved in DMSO at 10 mM and then diluted in culture medium at 0.01-10 µM final concentrations. Paclitaxel was purchased from Selleckchem (Houston, TX, USA).

Steinlein S., Essmann F., Ghilardi A.F., Horn H., Schüler J., Hausser A., Sun L., Ott G, & Kalla C. (2023). Indolyl-chalcone derivatives trigger apoptosis in cisplatin-resistant mesothelioma cells through aberrant tubulin polymerization and deregulation of microtubule-associated proteins. Frontiers in Oncology, 13, 1190988.

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