Anti phospho creb s133

Manufactured by Cell Signaling Technology

Anti-Phospho-CREB (S133) is a lab equipment product that detects the phosphorylated form of the CREB protein at serine 133. It is used for analysis of CREB phosphorylation in cell signaling studies.

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Close spelling variants of this product

Anti-CREB (111 citations) Phospho-CREB (67 citations) Anti-phospho-CREB (46 citations)

4 protocols using anti phospho creb s133

Immunoblotting of Cell Signaling Proteins

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The following antibodies and materials were used: anti-GAPDH (Calbiochem), anti-EGFR (Millipore), anti-CREB, anti-phospho CREB (S133), anti-STAT3, anti-phospho STAT3 (Y705), anti-c-Jun (Cell signaling), and anti-TGF-beta (Abcam). Electrophoresis reagents were purchased from Biorad.

Lachen-Montes M., Zelaya M.V., Segura V., Fernández-Irigoyen J, & Santamaría E. (2017). Progressive modulation of the human olfactory bulb transcriptome during Alzheimer´s disease evolution: novel insights into the olfactory signaling across proteinopathies. Oncotarget, 8(41), 69663-69679.

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Neuronal Signaling Pathways in PC12 Cells

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Cell media and chemicals, unless otherwise stated, were obtained from Sigma (St. Louis, MO). Fetal bovin serum (FBS), horse serum (HS), and NGF were obtained from Invitrogen Laboratories (Paisley, U.K.). NGF was used at a concentration of 50 ng/ml (approximately 4 × 10⁻⁹ M). Anti-phospho-TrkA (Y490) (Cat # 9141), anti-TrkA (Cat # 2505), anti-phospho-ERK1/2 (T202/Y204) (Cat # 9106), anti-ERK (Cat # 9107), anti-phospho-AKT (S473) (Cat # 4051), anti-AKT (Cat # 4685), anti-phospho-CREB (S133) (Cat # 9191), anti-CREB (Cat # 9197), and anti-P75^NTR (Cat # 4201) antibodies were from Cell Signaling Technology (Danvers, MA). Anti-Grb2 antibody (Cat # sc-17813) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rat pheochromocytoma (PC12) cells were obtained from the American Type Culture Collection (Manassas, VA), and cultured, at passages between the 5 and 20, in RPMI-1640 (GIBCO), supplemented with 10% horse serum (HS), 5% fetal bovine serum (FBS), 2 mM L-glutamine, 50 IU/ml penicillin, and 50 μg/ml streptomycin.

Pandini G., Satriano C., Pietropaolo A., Gianì F., Travaglia A., La Mendola D., Nicoletti V.G, & Rizzarelli E. (2016). The Inorganic Side of NGF: Copper(II) and Zinc(II) Affect the NGF Mimicking Signaling of the N-Terminus Peptides Encompassing the Recognition Domain of TrkA Receptor. Frontiers in Neuroscience, 10, 569.

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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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