Antibodies directed against AMPKα (1/1000, Rabbit), ACC (1/1000, Rabbit), phospho-Ser79ACC (1/1000, Rabbit), phospho-PKA substrate (RRXS*/T*) (1/2000, Rabbit), phospho-AMPK substrate (1/1000, Rabbit), and phospho-Ser133CREB (1/1000, Rabbit) were obtained from Cell Signaling technology (Danvers, MA, USA). Anti phospho-Thr172AMPKα (1/1000, Rabbit), CREB (1/500, Rabbit), Arc (1/500, Mouse), c-Fos (1/500, Mouse), and EgrI (1/500, Rabbit) antibodies were from Santa-Cruz (Dallas, TX, USA). Anti-actin (1/15 000, Mouse) antibody was from BD Bioscience (Franklin Lakes, NJ, USA). HRP-coupled secondary antibodies directed against the primary antibodies’ hosts were obtained from Cell Signaling technology. Bicuculline (Bic), H 89, PKI 14-22 amide, NKY 80, SQ 22536, and KH 7 were purchased from Tocris (Bristol, UK), 4-aminopyridine (4-AP) was purchased from Sigma (St Louis, MO, USA), and Compound C (Cc) was from Santa Cruz (Dallas, TX, USA).
Phospho ser133creb
Manufactured by Cell Signaling Technology
Sourced in United States
Phospho-Ser133CREB is a primary antibody that specifically recognizes the phosphorylated form of the transcription factor CREB (cAMP Response Element Binding protein) at serine 133. This post-translational modification plays a crucial role in the activation of CREB, which regulates the expression of genes involved in cellular processes such as memory formation, cell growth, and survival.
Automatically generated - may contain errors
Lab products found in correlation
Phospho jnk, by Cell Signaling Technology (2 mentions)
C fos, by Cell Signaling Technology (2 mentions)
Phospho erk, by Cell Signaling Technology (2 mentions)
Phospho p38, by Cell Signaling Technology (2 mentions)
Phospho thr71atf 2, by Cell Signaling Technology (2 mentions)
Phospho thr581 ser 376 ser360msk1, by Cell Signaling Technology (2 mentions)
β actin, by Merck Group (2 mentions)
Gapdh, by Santa Cruz Biotechnology (2 mentions)
C fos, by Santa Cruz Biotechnology (1 mentions)
Sq22536, by Bio-Techne (1 mentions)
Phospho acc ser79, by Cell Signaling Technology (1 mentions)
Phospho pka substrate rrxs t, by Cell Signaling Technology (1 mentions)
Phospho ampk substrate, by Cell Signaling Technology (1 mentions)
Pki 14 22 amide, by Bio-Techne (1 mentions)
Hrp coupled secondary antibody, by Cell Signaling Technology (1 mentions)
4 aminopyridine 4 ap, by Merck Group (1 mentions)
Anti actin, by BD (1 mentions)
Bovine pituitary extract, by Kurabo (1 mentions)
C jun, by Cell Signaling Technology (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
8 protocols using phospho ser133creb
1
Investigating AMPK-Mediated Signaling Pathways
2
Serum-free Keratinocyte Growth and Signaling
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Serum-free keratinocyte growth medium (Medium 154S) containing low calcium (0.2 mM), bovine pituitary extract (BPE) and epidermal growth factor (EGF) were obtained from Kurabo (Tokyo, Japan). Human primary keratinocytes (HPKs) were purchased from Biopredic Intl (Saint-Gregoire, France). Human HaCaT keratinocytes [22 (link), 23 (link)] were kindly supplied by Dr. M. Furue (Kyushu University, School of Medicine, Department of Dermatology). Rabbit polyclonal anti-TGase 1 (NOV-1184-45: NB100-1844 Lot7E3) was purchased from Novus Biologicals (Littleton, CO, USA). Antibodies to phospho-ERK, ERK, phospho-p38, p38, phospho-JNK, JNK, phospho-AKT and AKT were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies to phospho-Ser468/Ser276/Ser536NFκBp65, NFκBp65, phospho-Ser133CREB, CREB, c-Jun, phospho-Ser73c-Jun, c-Fos, ATF-2, phospho-Thr71ATF-2, MSK1 and phospho-Thr581/Ser 376/Ser360MSK1 were also purchased from Cell Signaling Technology. Antibodies to COX-2 and β-actin were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). The NFκB activation inhibitor II (JSH-23), the MEK inhibitor (PD98059), the p38 inhibitor (SB203580), the JNK inhibitor (JNK inhibitor II) and H89 were purchased from Calbiochem (San Diego, CA, USA). Other chemicals including AX were obtained from Sigma-Aldrich Corp.
3
Inhibition of PKCβ and Oxidative Stress
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PMA was purchased from Sigma-Aldrich (St. Louis, MO, USA). Anilino-monoindolylmaleimide (3-(1-(3-imidazol-1-ylpropyl)-1H-indol-3-yl)-4-anilino-1H-pyrrole-2,5-dione, a PKCβ-specific inhibitor (PKCβi), was purchased from Calbiochem (La Jolla, CA, USA). Triphenylphosphonium chloride (2-(2,2,6,6-tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) (Mito-TEMPO), a mitochondrial-specific antioxidant, was purchased from Enzo Life Sciences (Farmingdale, NY, USA). Specific antibodies against ICAM-1, GAPDH, PKCβII (Santa Cruz Biotechnology, CA), phospho-ser36-p66SHC (Calbiochem, CA), SHC (BD Biosciences, NJ), β-actin, Flag (Sigma, MO), phospho-ser133-CREB, CREB (Cell Signaling, MA), and MnSOD (Enzo Life Sciences, NY) were used in this study.
4
Westerns for Hippocampal Protein Analysis
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Protein was extracted from the hippocampi in rats with etomidate treatment or vehicle according to a previously described procedure.[14 (link)] Membranes were probed with primary antibodies against CREB, phospho-Ser133-CREB (#9104 and #9191, respectively, Cell Signaling, USA), Arc, c-fos, and Egr1 (ab118929, ab53036, and ab55160, respectively, Abcam, USA). Horseradish peroxidase conjugated anti-rabbit or anti-mouse secondary antibodies were used and visualized using the enhanced chemiluminescence kit (Thermo Scientific, USA). The density of each band was quantified using the ImageJ software (National Institutes of Health, USA). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ab1603, Abcam) was used as a loading control.
5
Signaling Pathway Antibodies and Inhibitors
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Antibodies to ERK, phospho-ERK, p38, phospho-p38, JNK, phospho-JNK, NFκBp65, phospho- Ser276/Ser536NFκBp65, CREB, phospho-Ser133CREB, c-Fos, ATF2, phospho-Thr71ATF2, MSK1 and phospho-Thr581/Ser 376/Ser360MSK1, JAK2 and phospho-Tyr221JAK2, STAT3 and phospho-Tyr705/Ser727STAT3 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies to β-actin and HYBID (Anti-KIAA1199) were obtained from Sigma-Aldrich Corp., St. Louis, MO, USA). The antibody to protein tyrosine phosphatase, nonreceptor type 9 (PTPMEG2/PTPN9) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The NF-κB activation inhibitor II (JSH-23), the MEK inhibitor (U0126), and the STAT3 inhibitor were from Calbiochem (San Diego, CA, USA). The p38 inhibitor (SB239063) and the JNK inhibitor (JNK inhibitor II) were from Santa Cruz Biotechnology and Merck KGaA (Darmstadt, Germany), respectively. Okadaic acid and sodium orthovanadate were obtained from Calbiochem.
6
Quercitrin Modulates Apoptosis Signaling
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DPCs (1 × 10 6 cells/dish) were seeded in 100 mm dishes and cultured for 24 h. Quercitrin was treated at concentrations of 1, 10, and 100 nM for appropriate time. The cells were then lysed and total cellular proteins were prepared. 50 μg protein samples were analyzed by Western blotting with corresponding antibodies; Bcl2 (1:1000, Abcam, cambridge, UK), Akt (1:1000, Santa Cruz, CA, USA), Erk (p44/42) (1:1000, Cell Signaling Technology, Danvers, MA, USA), CREB-1 (Cell Signaling Technology, Danvers, MA, USA) GAPDH (1:2000, Santa Cruz, CA, USA), phospho (Ser473)-Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA), phospho (Thr202/Tyr204)-Erk (p44/42) (1:1000, Cell Signaling Technology, Danvers, MA, USA), and phospho (Ser133)-CREB (1:1000, Cell Signaling Technology, Danvers, MA, USA). Western blot was analyzed by chemiluminescence detector iBright FL1000 (Invitrogen, Waltham, MA, USA).
7
Cell Lysis and Western Blotting
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For Western blot, cells were lysed in RIPA buffer with anti-protease and PhosSTOP tablets (Roche) and sonicated as previously described50 . Equal amounts of protein (20 μg) were resolved in a 4–12% Bis–Tris gel and transferred to a membrane using an iBLOT2 Dry Blotting System (Thermo Fisher Scientific). Membranes were immunoblotted with the following antibodies: mouse anti-SV40T (1/50; DP-02, Calbiochem Merck Biosciences), Phospho-Ser133 CREB (1:1,000; Cell Signaling Technology), CREB (1:1,000; Cell Signaling Technology), β-actin (1:2,000; Sigma), alpha-Tubulin (1:2,000; Sigma). Species-specific HRP-linked secondary antibodies (Cell Signaling Technology) were used for detection and visualization was performed on an ImageQuant LAS 4,000 following ECL exposure (GE Healthcare).
8
Immunoprecipitation of TPL-2 Kinase Signaling
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The 70-mer TPL-2 antibody used for immunoprecipitation of TPL-2 has been described previously [11 (link)]. Antibodies against MKK1/2, phospho(S217/S221)-MKK1/2, p38, phospho(T180/Y182)-p38, phospho-T573 RSK (p90 ribosomal S6 kinase), RSK1, phospho-S100 YB-1, YB-1, phospho(Ser133)-CREB (cyclic AMP response element-binding protein), CREB, phospho(Ser376)-MSK1, phospho(S189/S207)-MKK3/6, MKK6, MKK3, MKK4, phospho-MK2 (MAPKAP kinase 2), MK2, phospho(Ser257/Thr261)-MKK4, phospho(Ser271/Thr275)-MKK7, MKK7 JNK, phospho(T184) TAK1 (transforming growth factor-β-activated kinase 1), phospho(S176) IKK, phospho(S935) p105 and p105 were purchased from Cell Signaling Technology. Antibodies to TPL-2 (M20; H-7), Hsp90 and MKK4 were obtained from Santa Cruz. Phospho(T183/Y185)-JNK antibody was purchased from Invitrogen. Recombinant MBP-MKK6K82A and MKK1D208A proteins were obtained through the MRC PPU Reagents and Services facility (MRC PPU, College of Life Sciences, University of Dundee, UK).
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