Anti rsk1 rsk2 rsk3

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-RSK1/RSK2/RSK3 is a laboratory product that detects the presence and quantifies the levels of RSK1, RSK2, and RSK3 proteins in biological samples. This antibody-based tool can be used for various research applications that require the analysis of these kinases.

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4 protocols using anti rsk1 rsk2 rsk3

Validating Protein Targets via Western Blot

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To validate selected protein candidates, protein extracts from SKMEL24, A375 and the BRAFi-resistant sublines were analyzed by WB using NuPAGE Novex Bis-Tris Gel (Life Technologies) and PVDF membranes (Thermo Scientific), according to the manufacturer’s standard protocol. Antibodies for IHC and WB are purchased from: Anti-EPHA2 (sc-924) (IHC and WB) and anti-MET (sc-10) (IHC) SantaCruz Biotechnology INC.; Anti-EFNA1 (GTX63281) (WB) GeneTex; Anti-EFNA1 NBP1-30503 (IHC) and anti-FLI1 NBP1-95688 (IHC and WB) Novus Biologicals LLC (Littleton, CO, USA); Anti-phospho EPHA2 (Ser897) (WB) #6347, anti-MET (WB) #8198,, anti-pAKT1 (S129) #13461, anti-AKT1 #75692, anti-P-p90RSK (S380) #11989 and anti-RSK1/RSK2/RSK3 # 9355 Cell Signaling Technology (Danvers, MA, USA); anti-CD13/ANPEP #HPA004625 Protein Atlas; anti-β-actin (A5441) (WB) and anti-RRAS (WH0006237M1) (WB) Sigma life Science, Anti-CD13 (ab7417) (blocking antibody) Abcam, Cambridge, UK.

Azimi A., Tuominen R., Costa Svedman F., Caramuta S., Pernemalm M., Frostvik Stolt M., Kanter L., Kharaziha P., Lehtiö J., Hertzman Johansson C., Höiom V., Hansson J, & Egyhazi Brage S. (2017). Silencing FLI or targeting CD13/ANPEP lead to dephosphorylation of EPHA2, a mediator of BRAF inhibitor resistance, and induce growth arrest or apoptosis in melanoma cells. Cell Death & Disease, 8(8), e3029-.

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Comprehensive Immunoblot Analysis of mTOR Pathway

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Rabbit polyclonal anti-mTOR, anti-phosphorylated (p) mTOR^ser2448, anti-S6K1, anti-p-S6K1^thr389, anti-rS6, anti-p-rS6^ser235/236, anti-4EBP1, anti-p-4EBP1^ser65, anti-EIF4E, anti-p-EIF4E^ser209, anti-TSC1, anti-TSC2, anti-p-TSC2^ser1462, anti-p44/42 MAPK, anti-p-MAPK^{thr202/tyr204}, anti-RSK1/RSK2/RSK3, and anti-p-p90RSK^ser380 were obtained from Cell Signaling (Beverly, MA). Anti-vinculin antibody was purchased from Sigma Chemical (St. Louis, MO).

Calkins K.L., Thamotharan S., Dai Y., Shin B.C., Kalhan S.C, & Devaskar S.U. (2018). Early Dietary Restriction in Rats Alters Skeletal Muscle Tuberous Sclerosis Complex, Ribosomal s6 and Mitogen-Activated Protein Kinase. Nutrition research (New York, N.Y.), 54, 93-104.

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Western Blot Antibody Protocol

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Antibodies for Western blotting were used as follows: anti-O-GlcNAc (No. ab2739-Abcam): 1:1000; Anti-actin (No. A2066, Sigma Aldrich), 1:1000; Anti-Phospho-ERK1(T202/Y204)/ERK2(T185/Y187) (No. AF1018, R&D Systems), 1:500; Anti-ERK1/ERK2 (No. AF1576, R&D Systems), 1:500; Anti Phospho-p90RSK (S380) (No. 9341S, Cell Signaling), 1:500; Anti-Rsk1/rsk2/rsk3 (No. 9355T, Cell Signaling), 1:500; Anti-Phospho-Nurr77 (S351) (No. 5095S, Cell Signaling), 1:500; Anti-Nurr77 (No. NB100-56745, Novus Biologicals), 1:500; Anti-Phospho-CREB (S133) (No. 9198S, Cell Signaling), 1:500; Anti-CREB (No. 9104S, Cell Signaling), 1:500.

Massman L.J., Pereckas M., Zwagerman N.T, & Olivier-Van Stichelen S. (2021). O-GlcNAcylation Is Essential for Rapid Pomc Expression and Cell Proliferation in Corticotropic Tumor Cells. Endocrinology, 162(12), bqab178.

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Luciferase Assay for Pomc Promoter

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Antibodies for western blotting are used as follows: anti-O-GlcNAc (#ab2739-Abcam): 1:1000; Anti-actin (#A2066, Sigma Aldrich), 1:1000;
Anti-Phospho-ERK1(T202/Y204)/ERK2(T185/Y187) (#AF1018, R&D systems), 1:500; Anti-ERK1/ERK2 (#AF1576,R&D systems), 1:500; Anti Phospho-p90RSK (S380) (#9341S, Cell Signaling), 1:500; Anti-Rsk1/rsk2/rsk3 (#9355T, Cell Signaling), 1:500;
Anti-Phospho-Nurr77 (S351) (#5095S, Cell Signaling), 1:500; Anti-Nurr77 (#NB100-56745, Novus Biologicals), 1:500; Anti-Phospho-CREB (S133) (#9198S, Cell Signaling), 1:500; Anti-CREB (#9104S, Cell Signaling), 1:500.
AtT-20 cells were concurrently transfected using Lipofectamine 3000 (Thermofisher) with two plasmids -one containing the pRL Renilla luciferase gene under the control of the CMV promoter (# E2261, Promega) and the other containing the firefly luciferase gene under the control of the Pomc promoter (-646bp to +65bp) (#17553, Addgene). Forty-eight hours after transfection, luciferase activity was measured using the dual-luciferase assay protocol (#E1910, Promega). Transfection was performed 48h before measurement of the promoter activity.

Massman L., Pereckas M., Zwagerman N.T., & Stichelen S.O. (2021). O-GlcNAcylation is essential for rapid Pomc expression and cell proliferation in corticotropic tumor cells.

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