phospho creb  (Cell Signaling Technology Inc)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Name:
    Phospho CREB Ser133 87G3 Rabbit mAb
    Description:
    CREB is a bZIP transcription factor that activates target genes through cAMP response elements CREB is able to mediate signals from numerous physiological stimuli resulting in regulation of a broad array of cellular responses While CREB is expressed in numerous tissues it plays a large regulatory role in the nervous system CREB is believed to play a key role in promoting neuronal survival precursor proliferation neurite outgrowth and neuronal differentiation in certain neuronal populations 1 3 Additionally CREB signaling is involved in learning and memory in several organisms 4 6 CREB is able to selectively activate numerous downstream genes through interactions with different dimerization partners CREB is activated by phosphorylation at Ser133 by various signaling pathways including Erk Ca2 and stress signaling Some of the kinases involved in phosphorylating CREB at Ser133 are p90RSK MSK CaMKIV and MAPKAPK 2 7 9
    Catalog Number:
    9198
    Price:
    None
    Applications:
    Western Blot, Immunohistochemistry, Immunofluorescence, Flow Cytometry, Chromatin Immunoprecipitation
    Category:
    Primary Antibodies
    Source:
    Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser133 of human CREB.
    Reactivity:
    Human Mouse Rat
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc phospho creb
    Involvement of <t>MRP4</t> in regulation of embryo implantation gene expression and <t>CREB/COX-2</t> signaling (A) Quantitative PCR of COX-2, Claudin4, HoxA10, Lif and PPARg in mouse uteri 24 hours after intrauterine injection with MK-571 (0.5 mM, day 3 post mating) (n = 4, ** P
    CREB is a bZIP transcription factor that activates target genes through cAMP response elements CREB is able to mediate signals from numerous physiological stimuli resulting in regulation of a broad array of cellular responses While CREB is expressed in numerous tissues it plays a large regulatory role in the nervous system CREB is believed to play a key role in promoting neuronal survival precursor proliferation neurite outgrowth and neuronal differentiation in certain neuronal populations 1 3 Additionally CREB signaling is involved in learning and memory in several organisms 4 6 CREB is able to selectively activate numerous downstream genes through interactions with different dimerization partners CREB is activated by phosphorylation at Ser133 by various signaling pathways including Erk Ca2 and stress signaling Some of the kinases involved in phosphorylating CREB at Ser133 are p90RSK MSK CaMKIV and MAPKAPK 2 7 9
    https://www.bioz.com/result/phospho creb/product/Cell Signaling Technology Inc
    Average 90 stars, based on 92 article reviews
    Price from $9.99 to $1999.99
    phospho creb - by Bioz Stars, 2020-02
    90/100 stars

    Images

    1) Product Images from "MRP4 regulates ENaC-dependent CREB/COX-2/PGE2 signaling during embryo implantation"

    Article Title: MRP4 regulates ENaC-dependent CREB/COX-2/PGE2 signaling during embryo implantation

    Journal: Oncotarget

    doi: 10.18632/oncotarget.19676

    Involvement of MRP4 in regulation of embryo implantation gene expression and CREB/COX-2 signaling (A) Quantitative PCR of COX-2, Claudin4, HoxA10, Lif and PPARg in mouse uteri 24 hours after intrauterine injection with MK-571 (0.5 mM, day 3 post mating) (n = 4, ** P
    Figure Legend Snippet: Involvement of MRP4 in regulation of embryo implantation gene expression and CREB/COX-2 signaling (A) Quantitative PCR of COX-2, Claudin4, HoxA10, Lif and PPARg in mouse uteri 24 hours after intrauterine injection with MK-571 (0.5 mM, day 3 post mating) (n = 4, ** P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Injection

    2) Product Images from "Nanosecond pulsed electric fields enhanced chondrogenic potential of mesenchymal stem cells via JNK/CREB-STAT3 signaling pathway"

    Article Title: Nanosecond pulsed electric fields enhanced chondrogenic potential of mesenchymal stem cells via JNK/CREB-STAT3 signaling pathway

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-019-1133-0

    nsPEFs activated JNK, CREB, and STAT3 signaling pathways. a Western blot analysis for JNK, p-JNK with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). b Phosphorylation ratio for JNK with inhibitors, n = 5. c Western blot analysis for CREB, p-CREB with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). d Phosphorylation ratio for CREB with inhibitors, n = 5. e Western blot analysis for STAT3, p-STAT3 with inhibitors for phosphorylation of JNK (BI-78D3), CREB (BAPTA-AM), and STAT3 (Stattic). f Phosphorylation ratio for STAT3 with inhibitors, n = 5. Statistical significance in mean values was marked with different letters. For all variables with the same letter ( a , b , c , d , or e ), the difference between the groups is not statistically significant. For variables with a different letter, the difference between the groups is statistically significant ( P
    Figure Legend Snippet: nsPEFs activated JNK, CREB, and STAT3 signaling pathways. a Western blot analysis for JNK, p-JNK with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). b Phosphorylation ratio for JNK with inhibitors, n = 5. c Western blot analysis for CREB, p-CREB with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). d Phosphorylation ratio for CREB with inhibitors, n = 5. e Western blot analysis for STAT3, p-STAT3 with inhibitors for phosphorylation of JNK (BI-78D3), CREB (BAPTA-AM), and STAT3 (Stattic). f Phosphorylation ratio for STAT3 with inhibitors, n = 5. Statistical significance in mean values was marked with different letters. For all variables with the same letter ( a , b , c , d , or e ), the difference between the groups is not statistically significant. For variables with a different letter, the difference between the groups is statistically significant ( P

    Techniques Used: Western Blot

    nsPEFs promoted MSC chondrogenic differentiation through JNK/CREB-STAT3 signaling pathway. Expression levels for SOX9 , COL II , COL I , COL X , and ACAN in the absence or presence of inhibitors of either phosphorylation of CREB, JNK, or STAT3, or combination of them with ( a ) condition A, 10 ns at 20 kV/cm, and ( b ) condition B, 100 ns at 10 kV/cm. Diagonal (−) means inhibitors for corresponding proteins. Statistical significance in mean values was marked with different letters
    Figure Legend Snippet: nsPEFs promoted MSC chondrogenic differentiation through JNK/CREB-STAT3 signaling pathway. Expression levels for SOX9 , COL II , COL I , COL X , and ACAN in the absence or presence of inhibitors of either phosphorylation of CREB, JNK, or STAT3, or combination of them with ( a ) condition A, 10 ns at 20 kV/cm, and ( b ) condition B, 100 ns at 10 kV/cm. Diagonal (−) means inhibitors for corresponding proteins. Statistical significance in mean values was marked with different letters

    Techniques Used: Expressing

    3) Product Images from "Stem Cell Secretome and Its Effect on Cellular Mechanisms Relevant to Wound Healing"

    Article Title: Stem Cell Secretome and Its Effect on Cellular Mechanisms Relevant to Wound Healing

    Journal: Molecular Therapy

    doi: 10.1016/j.ymthe.2017.09.023

    The Stimulatory Effects of the Stem Cell Secretome on PI3K/Akt or FAK/ERK1/2 Signaling Serum-starved dermal cellular components (dermal fibroblasts, keratinocytes, and vascular endothelial cells) were stimulated for 30 min with either stem cell secretome or mock secretome (10 μg/mL). Cells were then lysed, and protein contents were analyzed by western blotting using antibodies targeting the phosphorylated forms of PI3K, Akt, and CREB (A) and FAK and ERK1/2 (B). The phosphorylation levels of these signaling molecules were significantly increased in cells treated with the stem cell secretome (A and B). β-actin was used as the internal control. The results are presented as the mean values ± SD from three independent experiments.
    Figure Legend Snippet: The Stimulatory Effects of the Stem Cell Secretome on PI3K/Akt or FAK/ERK1/2 Signaling Serum-starved dermal cellular components (dermal fibroblasts, keratinocytes, and vascular endothelial cells) were stimulated for 30 min with either stem cell secretome or mock secretome (10 μg/mL). Cells were then lysed, and protein contents were analyzed by western blotting using antibodies targeting the phosphorylated forms of PI3K, Akt, and CREB (A) and FAK and ERK1/2 (B). The phosphorylation levels of these signaling molecules were significantly increased in cells treated with the stem cell secretome (A and B). β-actin was used as the internal control. The results are presented as the mean values ± SD from three independent experiments.

    Techniques Used: Western Blot

    4) Product Images from "Rapamycin Blocks Induction of the Thermogenic Program in White Adipose Tissue"

    Article Title: Rapamycin Blocks Induction of the Thermogenic Program in White Adipose Tissue

    Journal: Diabetes

    doi: 10.2337/db15-0502

    Rapamycin (Rapa) attenuates beige fat gene expression in primary adipocytes. A and B : Expression of thermogenic genes, Ucp1 and Elovl3 , in primary adipocytes treated with 1 μmol/L CL ( A ) or 1 mmol/L 8-Br-cAMP (cAMP) ( B ) for 24 h with or without 500 nmol/L rapamycin pretreatment for 24 h. C and D : Adrb3 gene expression in the same cells treated with CL ( C ) or 8-Br-cAMP ( D ) with or without rapamycin pretreatment. E : PKA substrate phosphorylation (pPKA), phosphorylated CREB (pCREB), and pS6 in fully differentiated adipocytes treated with rapamycin for 24 h or PKA inhibitor for 1 h before treatment with CL for 20 min. Values shown are mean ± SEM. n = 4–6. * P
    Figure Legend Snippet: Rapamycin (Rapa) attenuates beige fat gene expression in primary adipocytes. A and B : Expression of thermogenic genes, Ucp1 and Elovl3 , in primary adipocytes treated with 1 μmol/L CL ( A ) or 1 mmol/L 8-Br-cAMP (cAMP) ( B ) for 24 h with or without 500 nmol/L rapamycin pretreatment for 24 h. C and D : Adrb3 gene expression in the same cells treated with CL ( C ) or 8-Br-cAMP ( D ) with or without rapamycin pretreatment. E : PKA substrate phosphorylation (pPKA), phosphorylated CREB (pCREB), and pS6 in fully differentiated adipocytes treated with rapamycin for 24 h or PKA inhibitor for 1 h before treatment with CL for 20 min. Values shown are mean ± SEM. n = 4–6. * P

    Techniques Used: Expressing

    5) Product Images from "Nanosecond pulsed electric fields enhanced chondrogenic potential of mesenchymal stem cells via JNK/CREB-STAT3 signaling pathway"

    Article Title: Nanosecond pulsed electric fields enhanced chondrogenic potential of mesenchymal stem cells via JNK/CREB-STAT3 signaling pathway

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-019-1133-0

    nsPEFs activated JNK, CREB, and STAT3 signaling pathways. a Western blot analysis for JNK, p-JNK with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). b Phosphorylation ratio for JNK with inhibitors, n = 5. c Western blot analysis for CREB, p-CREB with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). d Phosphorylation ratio for CREB with inhibitors, n = 5. e Western blot analysis for STAT3, p-STAT3 with inhibitors for phosphorylation of JNK (BI-78D3), CREB (BAPTA-AM), and STAT3 (Stattic). f Phosphorylation ratio for STAT3 with inhibitors, n = 5. Statistical significance in mean values was marked with different letters. For all variables with the same letter ( a , b , c , d , or e ), the difference between the groups is not statistically significant. For variables with a different letter, the difference between the groups is statistically significant ( P
    Figure Legend Snippet: nsPEFs activated JNK, CREB, and STAT3 signaling pathways. a Western blot analysis for JNK, p-JNK with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). b Phosphorylation ratio for JNK with inhibitors, n = 5. c Western blot analysis for CREB, p-CREB with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). d Phosphorylation ratio for CREB with inhibitors, n = 5. e Western blot analysis for STAT3, p-STAT3 with inhibitors for phosphorylation of JNK (BI-78D3), CREB (BAPTA-AM), and STAT3 (Stattic). f Phosphorylation ratio for STAT3 with inhibitors, n = 5. Statistical significance in mean values was marked with different letters. For all variables with the same letter ( a , b , c , d , or e ), the difference between the groups is not statistically significant. For variables with a different letter, the difference between the groups is statistically significant ( P

    Techniques Used: Western Blot

    nsPEFs promoted MSC chondrogenic differentiation through JNK/CREB-STAT3 signaling pathway. Expression levels for SOX9 , COL II , COL I , COL X , and ACAN in the absence or presence of inhibitors of either phosphorylation of CREB, JNK, or STAT3, or combination of them with ( a ) condition A, 10 ns at 20 kV/cm, and ( b ) condition B, 100 ns at 10 kV/cm. Diagonal (−) means inhibitors for corresponding proteins. Statistical significance in mean values was marked with different letters
    Figure Legend Snippet: nsPEFs promoted MSC chondrogenic differentiation through JNK/CREB-STAT3 signaling pathway. Expression levels for SOX9 , COL II , COL I , COL X , and ACAN in the absence or presence of inhibitors of either phosphorylation of CREB, JNK, or STAT3, or combination of them with ( a ) condition A, 10 ns at 20 kV/cm, and ( b ) condition B, 100 ns at 10 kV/cm. Diagonal (−) means inhibitors for corresponding proteins. Statistical significance in mean values was marked with different letters

    Techniques Used: Expressing

    Related Articles

    Centrifugation:

    Article Title: TNFα-signal and cAMP-mediated signals oppositely regulate melanoma- associated ganglioside GD3 synthase gene in human melanocytes
    Article Snippet: Insoluble materials were removed by centrifugation at 10,000 × g for 10 min at 4°C. .. Monoclonal antibodies used for detection of CREB and phospho-CREB were as follows, CREB (48H2) Rabbit mAb (Cell Signaling Technology); Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology).

    Article Title: Novel GLP-1R/GIPR co-agonist “twincretin” is neuroprotective in cell and rodent models of mild traumatic brain injury
    Article Snippet: After light centrifugation, the DPBS supernatant was removed, and the cells were lysed with a 100:1 M-PER Mammalian Protein Extraction Reagent (ThermoFisher Scientific, Waltham, MA), Halt Protease and Phosphatase Inhibitor Cocktail (ThermoFisher Scientific, Waltham, MA) mixture. .. The three primary antibodies used in this study were Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology, Danvers, MA), CREB (48H2) Rabbit mAb (Cell Signaling Technology, Danvers, MA) and α-tubulin Mouse mAb (Sigma-Aldrich, St. Louis, MO).

    Immunocytochemistry:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: Paragraph title: Histology, immunohistochemistry (IHC) and immunocytochemistry (ICC) ... For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana).

    MTT Assay:

    Article Title: Involvement of Akt/CREB signaling pathways in the protective effect of EPA against interleukin-1β-induced cytotoxicity and BDNF down-regulation in cultured rat hippocampal neurons
    Article Snippet: 3-(4,5-Dimethylthiazol-2-yl)-2,5-dimethyltetrazolium bromide (MTT), DNAase1 and 1-β-D-arabinofuranosylcytosine were obtained from Sigma-Aldrich (Saint Louis, MO, USA). .. Akt (catalog No. 9272), phospho-Akt (catalog No. 5012S, Ser473), CREB (catalog No. 4820) and phospho-CREB (catalog No. 9198, Ser133) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Article Title: Antiphotoaging and Antimelanogenic Effects of Penthorum chinense Pursh Ethanol Extract due to Antioxidant- and Autophagy-Inducing Properties
    Article Snippet: 3-(4-5-Dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide (MTT), 5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one (kojic acid), monophenol monooxygenase (mushroom tyrosinase), 4-hydroxyphenyl-β -D-glucopyranoside (arbutin), 1,1-diphenyl-2-picrylhydrazyl (DPPH), α -MSH, L-DOPA ethyl ester, ascorbic acid, and forskolin were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. Total and phospho-specific antibodies of CREB (#9198), phospho-CREB (#9198), lamin A/C (#4777), β -actin (#4967), ERK (#4696), phospho-ERK (#9101), p38 (#9212), phospho-p38 (#4631), JNK (#4672), phospho-JNK (#9255), and LC3B (#2775) were purchased from Cell Signaling Technology (Beverly, MA, USA).

    Blocking Assay:

    Article Title: Phosphodiesterase Inhibition Increases CREB Phosphorylation and Restores Orientation Selectivity in a Model of Fetal Alcohol Spectrum Disorders
    Article Snippet: .. Blots were incubated in blocking buffer (LI-COR, Lincoln, NE) for 1 hour, then incubated in Phospho-CREB (Ser133) (87G3) rabbit mAb (1∶200, Cell Signalling Tech.) overnight at 4°C. .. This was followed by two hours of incubation in CREB (86B10) mouse mAb (1∶200, Cell Signalling Tech.) at room temperature, then one hour in GAPDH mouse mAb (Sigma, St. Louis, MO).

    Article Title: Activation of PTHrP-cAMP-CREB1 signaling following p53 loss is essential for osteosarcoma initiation and maintenance
    Article Snippet: All antibodies (p53 (1C12) Cell Signaling 2524) (Phospho-CREB (Ser133) Cell Signaling Technologies, #9198), Anti-CREB1 ChIP grade (ab31387) were used at 1:2000, except pan actin (Ab-5, Thermo Scientific, Waltham, MA, USA) that was used at 1:3000. .. Protocol for PTHrP western is the same as described for CREB1 except 3% BSA was used for blocking instead of skim milk.

    SYBR Green Assay:

    Article Title: Involvement of Akt/CREB signaling pathways in the protective effect of EPA against interleukin-1β-induced cytotoxicity and BDNF down-regulation in cultured rat hippocampal neurons
    Article Snippet: Akt (catalog No. 9272), phospho-Akt (catalog No. 5012S, Ser473), CREB (catalog No. 4820) and phospho-CREB (catalog No. 9198, Ser133) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). .. An Ominiscript RT Kit and SYBR Green Kit were purchased from QIAGEN (Hilden, Germany).

    Incubation:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: After incubation for 1 h with a secondary Alexa488-linked anti-rabbit polyclonal antibody (Cell Signaling) at 4°C, the cells were washed with HBSS-T following HBSS. .. For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana).

    Article Title: Phosphodiesterase Inhibition Increases CREB Phosphorylation and Restores Orientation Selectivity in a Model of Fetal Alcohol Spectrum Disorders
    Article Snippet: .. Blots were incubated in blocking buffer (LI-COR, Lincoln, NE) for 1 hour, then incubated in Phospho-CREB (Ser133) (87G3) rabbit mAb (1∶200, Cell Signalling Tech.) overnight at 4°C. .. This was followed by two hours of incubation in CREB (86B10) mouse mAb (1∶200, Cell Signalling Tech.) at room temperature, then one hour in GAPDH mouse mAb (Sigma, St. Louis, MO).

    Article Title: TNFα-signal and cAMP-mediated signals oppositely regulate melanoma- associated ganglioside GD3 synthase gene in human melanocytes
    Article Snippet: Blots were blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 or 3% BSA in PBS containing 0.05% Tween 20 for 1 h at room temperature, and incubated with anti-IKKα, anti-IKKβ antibody, or anti-Phospho-IKKα/β (Ser176/180) antibody (Cell Signaling Technology) as primary antibodies. .. Monoclonal antibodies used for detection of CREB and phospho-CREB were as follows, CREB (48H2) Rabbit mAb (Cell Signaling Technology); Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology).

    Article Title: Altered phosphorylation, electrophysiology, and behavior on attenuation of PDE4B action in hippocampus
    Article Snippet: One half of the 5th series was stained for phospho-CREB (pCREB, rabbit, Cell Signaling, #9198), whereas the other half was stained for CREB (rabbit, Cell Signaling, #9197). .. Following incubation in this solution for 18 h on a shaker table at room temperature (20 °C) in the dark, the sections were rinsed 3 times in TBS-T and transferred to the solution containing the appropriate secondary antibody (goat anti-mouse*biotin or donkey anti-rabbit*biotin, Sigma).

    Article Title: Activation of PTHrP-cAMP-CREB1 signaling following p53 loss is essential for osteosarcoma initiation and maintenance
    Article Snippet: Membranes were blocked in 5% skim milk in TBST (20 mM Tris, 150 mM NaCl, 0.1% Tween-20) for 1 hr before incubation with primary antibodies diluted in 5% skim milk in TBST overnight a 4°C, or for 1 hr at room temperature in the case of pan-ACTIN. .. All antibodies (p53 (1C12) Cell Signaling 2524) (Phospho-CREB (Ser133) Cell Signaling Technologies, #9198), Anti-CREB1 ChIP grade (ab31387) were used at 1:2000, except pan actin (Ab-5, Thermo Scientific, Waltham, MA, USA) that was used at 1:3000.

    Article Title: Novel GLP-1R/GIPR co-agonist “twincretin” is neuroprotective in cell and rodent models of mild traumatic brain injury
    Article Snippet: The membrane was then blocked with a 5% milk solution in tris-buffered saline and Tween 20 (TBST) for 1 h prior to overnight incubation with primary antibody in 5% milk solution in a cold room. .. The three primary antibodies used in this study were Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology, Danvers, MA), CREB (48H2) Rabbit mAb (Cell Signaling Technology, Danvers, MA) and α-tubulin Mouse mAb (Sigma-Aldrich, St. Louis, MO).

    Luciferase:

    Article Title: Antiphotoaging and Antimelanogenic Effects of Penthorum chinense Pursh Ethanol Extract due to Antioxidant- and Autophagy-Inducing Properties
    Article Snippet: Luciferase plasmids harboring promoter binding sites for CREB and Col1A1 were used as reported earlier [ ]. .. Total and phospho-specific antibodies of CREB (#9198), phospho-CREB (#9198), lamin A/C (#4777), β -actin (#4967), ERK (#4696), phospho-ERK (#9101), p38 (#9212), phospho-p38 (#4631), JNK (#4672), phospho-JNK (#9255), and LC3B (#2775) were purchased from Cell Signaling Technology (Beverly, MA, USA).

    BIA-KA:

    Article Title: Involvement of Akt/CREB signaling pathways in the protective effect of EPA against interleukin-1β-induced cytotoxicity and BDNF down-regulation in cultured rat hippocampal neurons
    Article Snippet: The protease inhibitor mixture, phosphatase inhibitor, BCA protein assay kit and enhanced chemiluminescence substrate kit were obtained from Pierce Biotechnology (Rockford, IL, USA). .. Akt (catalog No. 9272), phospho-Akt (catalog No. 5012S, Ser473), CREB (catalog No. 4820) and phospho-CREB (catalog No. 9198, Ser133) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Article Title: Ketogenic diet improves the spatial memory impairment caused by exposure to hypobaric hypoxia through increased acetylation of histones in rats
    Article Snippet: The protein concentration was determined using a Pierce BCA Protein Assay kit (Thermo Scientific). .. A quantity of 50 μg of total protein was loaded onto a 10–15% SDS-PAGE, electrophoretically transferred to polyvinylidene difluoride membrane (PVDF), and probed with the following primary antibodies: Acetyl-Histone H3 (06–599, Merck Millipore), Acetyl-Histone H3 (Lys14) (07–353, Merck Millipore), Histone H4 (Acetyl K12) (ab61238, Abcam), Histone H3 (4499, Cell Signaling Tech.), Histone H4 (2935, Cell Signaling Tech.), phospho-CREB (Ser133) (9198, Cell Signaling Tech.), CREB (4820, Cell Signaling Tech.), phospho-(Ser/Thr) PKA substrates (9621, Cell Signaling Tech). β-actin (A2228, Sigma-Aldrich) was used as an internal control.

    Article Title: Novel GLP-1R/GIPR co-agonist “twincretin” is neuroprotective in cell and rodent models of mild traumatic brain injury
    Article Snippet: The total protein content in each sample was determined via BCA assay (ThermoFisher Scientific, Waltham, MA). .. The three primary antibodies used in this study were Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology, Danvers, MA), CREB (48H2) Rabbit mAb (Cell Signaling Technology, Danvers, MA) and α-tubulin Mouse mAb (Sigma-Aldrich, St. Louis, MO).

    Modification:

    Article Title: Antiphotoaging and Antimelanogenic Effects of Penthorum chinense Pursh Ethanol Extract due to Antioxidant- and Autophagy-Inducing Properties
    Article Snippet: Fetal bovine serum and Dulbecco's modified Eagle's media (DMEM) were purchased from Gibco (Grand Island, NY, USA). .. Total and phospho-specific antibodies of CREB (#9198), phospho-CREB (#9198), lamin A/C (#4777), β -actin (#4967), ERK (#4696), phospho-ERK (#9101), p38 (#9212), phospho-p38 (#4631), JNK (#4672), phospho-JNK (#9255), and LC3B (#2775) were purchased from Cell Signaling Technology (Beverly, MA, USA).

    Western Blot:

    Article Title: Phosphodiesterase Inhibition Increases CREB Phosphorylation and Restores Orientation Selectivity in a Model of Fetal Alcohol Spectrum Disorders
    Article Snippet: Paragraph title: Western blotting ... Blots were incubated in blocking buffer (LI-COR, Lincoln, NE) for 1 hour, then incubated in Phospho-CREB (Ser133) (87G3) rabbit mAb (1∶200, Cell Signalling Tech.) overnight at 4°C.

    Article Title: TNFα-signal and cAMP-mediated signals oppositely regulate melanoma- associated ganglioside GD3 synthase gene in human melanocytes
    Article Snippet: Paragraph title: Western immunoblotting ... Monoclonal antibodies used for detection of CREB and phospho-CREB were as follows, CREB (48H2) Rabbit mAb (Cell Signaling Technology); Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology).

    Article Title: Activation of PTHrP-cAMP-CREB1 signaling following p53 loss is essential for osteosarcoma initiation and maintenance
    Article Snippet: Paragraph title: Western blotting ... All antibodies (p53 (1C12) Cell Signaling 2524) (Phospho-CREB (Ser133) Cell Signaling Technologies, #9198), Anti-CREB1 ChIP grade (ab31387) were used at 1:2000, except pan actin (Ab-5, Thermo Scientific, Waltham, MA, USA) that was used at 1:3000.

    Article Title: Ketogenic diet improves the spatial memory impairment caused by exposure to hypobaric hypoxia through increased acetylation of histones in rats
    Article Snippet: Paragraph title: Western blot analysis ... A quantity of 50 μg of total protein was loaded onto a 10–15% SDS-PAGE, electrophoretically transferred to polyvinylidene difluoride membrane (PVDF), and probed with the following primary antibodies: Acetyl-Histone H3 (06–599, Merck Millipore), Acetyl-Histone H3 (Lys14) (07–353, Merck Millipore), Histone H4 (Acetyl K12) (ab61238, Abcam), Histone H3 (4499, Cell Signaling Tech.), Histone H4 (2935, Cell Signaling Tech.), phospho-CREB (Ser133) (9198, Cell Signaling Tech.), CREB (4820, Cell Signaling Tech.), phospho-(Ser/Thr) PKA substrates (9621, Cell Signaling Tech). β-actin (A2228, Sigma-Aldrich) was used as an internal control.

    Article Title: Novel GLP-1R/GIPR co-agonist “twincretin” is neuroprotective in cell and rodent models of mild traumatic brain injury
    Article Snippet: Paragraph title: 2.4. Western blotting ... The three primary antibodies used in this study were Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology, Danvers, MA), CREB (48H2) Rabbit mAb (Cell Signaling Technology, Danvers, MA) and α-tubulin Mouse mAb (Sigma-Aldrich, St. Louis, MO).

    Flow Cytometry:

    Article Title: Activation of PTHrP-cAMP-CREB1 signaling following p53 loss is essential for osteosarcoma initiation and maintenance
    Article Snippet: All antibodies (p53 (1C12) Cell Signaling 2524) (Phospho-CREB (Ser133) Cell Signaling Technologies, #9198), Anti-CREB1 ChIP grade (ab31387) were used at 1:2000, except pan actin (Ab-5, Thermo Scientific, Waltham, MA, USA) that was used at 1:3000. .. IgG was extracted from whole serum using Protein G Sepharose 4B fast Flow (GE Healthcare Life Sciences, Cat number 17-0618-01).

    Protease Inhibitor:

    Article Title: Involvement of Akt/CREB signaling pathways in the protective effect of EPA against interleukin-1β-induced cytotoxicity and BDNF down-regulation in cultured rat hippocampal neurons
    Article Snippet: The protease inhibitor mixture, phosphatase inhibitor, BCA protein assay kit and enhanced chemiluminescence substrate kit were obtained from Pierce Biotechnology (Rockford, IL, USA). .. Akt (catalog No. 9272), phospho-Akt (catalog No. 5012S, Ser473), CREB (catalog No. 4820) and phospho-CREB (catalog No. 9198, Ser133) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Generated:

    Article Title: Activation of PTHrP-cAMP-CREB1 signaling following p53 loss is essential for osteosarcoma initiation and maintenance
    Article Snippet: All antibodies (p53 (1C12) Cell Signaling 2524) (Phospho-CREB (Ser133) Cell Signaling Technologies, #9198), Anti-CREB1 ChIP grade (ab31387) were used at 1:2000, except pan actin (Ab-5, Thermo Scientific, Waltham, MA, USA) that was used at 1:3000. .. Anti-PTHrP antibody (R88) was generated in house against PTHrP(1–15).

    other:

    Article Title: Corticotropin releasing hormone receptor 2 exacerbates chronic cardiac dysfunction
    Article Snippet: Materials and chemicals Antibodies to Crhr2 (sc-20550), β-actin (sc-47778), Adrb1 (sc-568), Ptger1 (sc-22648), CaMKII (sc-5306), phospho-CaMKII (Thr286, sc-32289), and RyR (sc-376507) were obtained from Santa Cruz Biotechnology, Inc. Antibodies to CREB (48H2, #9197), phospho-CREB (Ser133, 87G3, #9198), PKA (#4782), phospho-PKA (Thr197, #4781), AKT (#9272), and phospho-AKT (Ser473, #4060) were all from Cell Signaling Technology.

    Article Title: A Dual Array-Based Approach to Assess the Abundance and Posttranslational Modification State of Signaling Proteins
    Article Snippet: Anti–β-actin antibody (Sigma, #A1978) Antibody against Akt (protein kinase B) phosphorylated at Ser473 (Cell Signaling Technology, #4058) Anti-axin1 antibody (Cell Signaling Technology, #2087) Anti-axin2 antibody (Cell Signaling Technology, #2151) Anti–β-catenin antibody (Cell Signaling Technology, #9582) Antibody against β-catenin phosphorylated at Ser675 (Cell Signaling Technology, #9567) Anti–casein kinase 1 (CK1) antibody (Cell Signaling Technology, #2655) Anti–casein kinase 2α (CK2α) antibody (Cell Signaling Technology, #2656) Antibody against cAMP response element-binding (CREB) phosphorylated at Ser133 (Cell Signaling Technology, #9198) Anti–Dishevelled 2 (Dvl2) antibody (Cell Signaling Technology, #3224) Anti–Dishevelled 3 (Dvl3) antibody (Cell Signaling Technology, #3218) Antibody against extracellular signal–regulated kinases (ERK1/2) phopshorylated at (Thr202 /Tyr204 ) (Cell Signaling Technology, #4377) Anti–low-density lipoprotein receptor–related protein 6 (LRP6) antibody (Cell Signaling Technology, #2560) Antibody against LRP6 phosphorylated at Ser1490 (Cell Signaling Technology, #2568) Antibody against mitogen-activated protein kinase kinase 1/2 (MEK1/2) phosphorylated at Ser217/221 (Cell Signaling Technology, #9121) Antibody against protein kinase C (PKC) phosphorylated at Thr410 (Cell Signaling Technology, #2060) Antibody against Raf phosphorylated at Ser289/296/301 (Cell Signaling Technology, #9431) Antibody against signal transducers and activators of transcription (STAT) 1 phosphorylated at Tyr701 (Cell Signaling Technology, #9167) Antibody against STAT3 phosphorylated at Tyr705 (Cell Signaling Technology, #9145) Anti–TCF1 antibody (Cell Signaling Technology, #2203) Anti-TCF3 antibody (Cell Signaling Technology, #2883)

    Article Title: Xiaoyao Pills Prevent Lipopolysaccharide-Induced Depression by Inhibiting Inflammation and Protecting Nerves
    Article Snippet: Rabbit anti-TrkB antibody (1:1000; Cell Signaling Technology; Cat. No. #4603), rabbit anti-cAMP response element-binding protein (CREB) antibody (1:1000; Cell Signaling Technology; Cat. No. #9197S), rabbit anti-p-CREB antibody (1:1000; Cell Signaling Technology; Cat. No. #9198S), rabbit anti-β-Tubulin antibody (1:1000; Cell Signaling Technology; Cat. No. #2128).

    Protein Concentration:

    Article Title: Ketogenic diet improves the spatial memory impairment caused by exposure to hypobaric hypoxia through increased acetylation of histones in rats
    Article Snippet: The protein concentration was determined using a Pierce BCA Protein Assay kit (Thermo Scientific). .. A quantity of 50 μg of total protein was loaded onto a 10–15% SDS-PAGE, electrophoretically transferred to polyvinylidene difluoride membrane (PVDF), and probed with the following primary antibodies: Acetyl-Histone H3 (06–599, Merck Millipore), Acetyl-Histone H3 (Lys14) (07–353, Merck Millipore), Histone H4 (Acetyl K12) (ab61238, Abcam), Histone H3 (4499, Cell Signaling Tech.), Histone H4 (2935, Cell Signaling Tech.), phospho-CREB (Ser133) (9198, Cell Signaling Tech.), CREB (4820, Cell Signaling Tech.), phospho-(Ser/Thr) PKA substrates (9621, Cell Signaling Tech). β-actin (A2228, Sigma-Aldrich) was used as an internal control.

    Binding Assay:

    Article Title: Antiphotoaging and Antimelanogenic Effects of Penthorum chinense Pursh Ethanol Extract due to Antioxidant- and Autophagy-Inducing Properties
    Article Snippet: Luciferase plasmids harboring promoter binding sites for CREB and Col1A1 were used as reported earlier [ ]. .. Total and phospho-specific antibodies of CREB (#9198), phospho-CREB (#9198), lamin A/C (#4777), β -actin (#4967), ERK (#4696), phospho-ERK (#9101), p38 (#9212), phospho-p38 (#4631), JNK (#4672), phospho-JNK (#9255), and LC3B (#2775) were purchased from Cell Signaling Technology (Beverly, MA, USA).

    Nucleic Acid Electrophoresis:

    Article Title: MRP4 regulates ENaC-dependent CREB/COX-2/PGE2 signaling during embryo implantation
    Article Snippet: Equal amounts of protein were resolved by SDS-polyacrylamide gel electrophoresis and electroblotted onto equilibrated nitrocellulose membrane. .. Antibodies against MRP4 (1:100, Abcam, ab15602); phospho-CREB (1:1000, Cell Signaling, 9198), β-tubulin (1:2000, Santa Cruz, sc-9104) and β-actin (1:5000, Sigma, A1978).

    Fluorescence:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: DAPI was employed for the nuclear staining and fluorescence was analysed with a fluorescence microscope (Leica). .. For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana).

    Immunohistochemistry:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: Paragraph title: Histology, immunohistochemistry (IHC) and immunocytochemistry (ICC) ... For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana).

    Article Title: Altered phosphorylation, electrophysiology, and behavior on attenuation of PDE4B action in hippocampus
    Article Snippet: Paragraph title: Immunohistochemistry ... One half of the 5th series was stained for phospho-CREB (pCREB, rabbit, Cell Signaling, #9198), whereas the other half was stained for CREB (rabbit, Cell Signaling, #9197).

    Microscopy:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: DAPI was employed for the nuclear staining and fluorescence was analysed with a fluorescence microscope (Leica). .. For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana).

    Mouse Assay:

    Article Title: Altered phosphorylation, electrophysiology, and behavior on attenuation of PDE4B action in hippocampus
    Article Snippet: Immunohistochemistry Antibody staining in the CNS of transgenic and wild-type mice (2–3 mice in each experimental group, or as specified in Results) was performed as described previously [ ]. .. One half of the 5th series was stained for phospho-CREB (pCREB, rabbit, Cell Signaling, #9198), whereas the other half was stained for CREB (rabbit, Cell Signaling, #9197).

    Transgenic Assay:

    Article Title: Altered phosphorylation, electrophysiology, and behavior on attenuation of PDE4B action in hippocampus
    Article Snippet: Immunohistochemistry Antibody staining in the CNS of transgenic and wild-type mice (2–3 mice in each experimental group, or as specified in Results) was performed as described previously [ ]. .. One half of the 5th series was stained for phospho-CREB (pCREB, rabbit, Cell Signaling, #9198), whereas the other half was stained for CREB (rabbit, Cell Signaling, #9197).

    Protein Extraction:

    Article Title: Novel GLP-1R/GIPR co-agonist “twincretin” is neuroprotective in cell and rodent models of mild traumatic brain injury
    Article Snippet: After light centrifugation, the DPBS supernatant was removed, and the cells were lysed with a 100:1 M-PER Mammalian Protein Extraction Reagent (ThermoFisher Scientific, Waltham, MA), Halt Protease and Phosphatase Inhibitor Cocktail (ThermoFisher Scientific, Waltham, MA) mixture. .. The three primary antibodies used in this study were Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology, Danvers, MA), CREB (48H2) Rabbit mAb (Cell Signaling Technology, Danvers, MA) and α-tubulin Mouse mAb (Sigma-Aldrich, St. Louis, MO).

    Bradford Protein Assay:

    Article Title: Phosphodiesterase Inhibition Increases CREB Phosphorylation and Restores Orientation Selectivity in a Model of Fetal Alcohol Spectrum Disorders
    Article Snippet: Protein concentrations were measured using Bradford protein assay with bovine serum albumin as a standard (Bio-Rad, Hercules, CA). .. Blots were incubated in blocking buffer (LI-COR, Lincoln, NE) for 1 hour, then incubated in Phospho-CREB (Ser133) (87G3) rabbit mAb (1∶200, Cell Signalling Tech.) overnight at 4°C.

    Staining:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: .. For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana). .. Determination of the MMP activity by zymography Gelatin degradation activity of MMP-2 and 9 in the culture supernatant was determined with the gelatin zymography as recently described [ ].

    Article Title: Altered phosphorylation, electrophysiology, and behavior on attenuation of PDE4B action in hippocampus
    Article Snippet: .. One half of the 5th series was stained for phospho-CREB (pCREB, rabbit, Cell Signaling, #9198), whereas the other half was stained for CREB (rabbit, Cell Signaling, #9197). .. One half of the 6th series was stained for phospho-ERK1/2 (pERK1/2, pT202/pY204, rabbit, Cell Signaling, #9101), whereas the other half was stained for ERK1/2 (rabbit, Cell Signaling, #9102).

    Article Title: Novel GLP-1R/GIPR co-agonist “twincretin” is neuroprotective in cell and rodent models of mild traumatic brain injury
    Article Snippet: The three primary antibodies used in this study were Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology, Danvers, MA), CREB (48H2) Rabbit mAb (Cell Signaling Technology, Danvers, MA) and α-tubulin Mouse mAb (Sigma-Aldrich, St. Louis, MO). .. For the membranes stained for pCREB and CREB, 1:5000 dilutions of HRP-conjugated goat anti-rabbit antibody (sc-2030, Santa Cruz Biotechnology, Dallas, TX) were used.

    Chromatin Immunoprecipitation:

    Article Title: Genes that Confer the Identity of the Renin Cell
    Article Snippet: Paragraph title: Chromatin Immunoprecipitation (ChIP) ... The antibodies used were Phospho-Creb (Ser 133, 9198S; Cell Signaling Technology, Inc., Danvers, MA) and RBP-J (AB5790; Millipore, Billerica, MA).

    Article Title: Activation of PTHrP-cAMP-CREB1 signaling following p53 loss is essential for osteosarcoma initiation and maintenance
    Article Snippet: .. All antibodies (p53 (1C12) Cell Signaling 2524) (Phospho-CREB (Ser133) Cell Signaling Technologies, #9198), Anti-CREB1 ChIP grade (ab31387) were used at 1:2000, except pan actin (Ab-5, Thermo Scientific, Waltham, MA, USA) that was used at 1:3000. .. Anti-PTHrP antibody (R88) was generated in house against PTHrP(1–15).

    SDS Page:

    Article Title: Phosphodiesterase Inhibition Increases CREB Phosphorylation and Restores Orientation Selectivity in a Model of Fetal Alcohol Spectrum Disorders
    Article Snippet: Next, 50 µg of total protein was resolved by SDS-PAGE (10% Tris-HCl Rgels, Bio-Rad, Hercules, CA) and transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA). .. Blots were incubated in blocking buffer (LI-COR, Lincoln, NE) for 1 hour, then incubated in Phospho-CREB (Ser133) (87G3) rabbit mAb (1∶200, Cell Signalling Tech.) overnight at 4°C.

    Article Title: TNFα-signal and cAMP-mediated signals oppositely regulate melanoma- associated ganglioside GD3 synthase gene in human melanocytes
    Article Snippet: Cell lysates were separated by SDS-PAGE using 10% gels. .. Monoclonal antibodies used for detection of CREB and phospho-CREB were as follows, CREB (48H2) Rabbit mAb (Cell Signaling Technology); Phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling Technology).

    Article Title: Ketogenic diet improves the spatial memory impairment caused by exposure to hypobaric hypoxia through increased acetylation of histones in rats
    Article Snippet: .. A quantity of 50 μg of total protein was loaded onto a 10–15% SDS-PAGE, electrophoretically transferred to polyvinylidene difluoride membrane (PVDF), and probed with the following primary antibodies: Acetyl-Histone H3 (06–599, Merck Millipore), Acetyl-Histone H3 (Lys14) (07–353, Merck Millipore), Histone H4 (Acetyl K12) (ab61238, Abcam), Histone H3 (4499, Cell Signaling Tech.), Histone H4 (2935, Cell Signaling Tech.), phospho-CREB (Ser133) (9198, Cell Signaling Tech.), CREB (4820, Cell Signaling Tech.), phospho-(Ser/Thr) PKA substrates (9621, Cell Signaling Tech). β-actin (A2228, Sigma-Aldrich) was used as an internal control. .. The membranes were developed using an enhanced chemiluminescence detection system (Pierce, Rockford, IL).

    Software:

    Article Title: Ketogenic diet improves the spatial memory impairment caused by exposure to hypobaric hypoxia through increased acetylation of histones in rats
    Article Snippet: A quantity of 50 μg of total protein was loaded onto a 10–15% SDS-PAGE, electrophoretically transferred to polyvinylidene difluoride membrane (PVDF), and probed with the following primary antibodies: Acetyl-Histone H3 (06–599, Merck Millipore), Acetyl-Histone H3 (Lys14) (07–353, Merck Millipore), Histone H4 (Acetyl K12) (ab61238, Abcam), Histone H3 (4499, Cell Signaling Tech.), Histone H4 (2935, Cell Signaling Tech.), phospho-CREB (Ser133) (9198, Cell Signaling Tech.), CREB (4820, Cell Signaling Tech.), phospho-(Ser/Thr) PKA substrates (9621, Cell Signaling Tech). β-actin (A2228, Sigma-Aldrich) was used as an internal control. .. For densitometry of the protein bands, the optical densities (OD) of the protein bands were quantified using Quantity One software (Bio-Rad).

    Concentration Assay:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: .. For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana). .. Determination of the MMP activity by zymography Gelatin degradation activity of MMP-2 and 9 in the culture supernatant was determined with the gelatin zymography as recently described [ ].

    Formalin-fixed Paraffin-Embedded:

    Article Title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization
    Article Snippet: .. For evaluation of localization of CREB in human breast cancer tissue, formalin-fixed paraffin embedded tissue samples that contained necrotic areas indicative of localized hypoxia were stained with the phospho-CREB (Ser133) (87G3) Rabbit mAb (Cell Signaling) in the indicated concentration using the Ventana Discovery Autostainer (Ventana). .. Determination of the MMP activity by zymography Gelatin degradation activity of MMP-2 and 9 in the culture supernatant was determined with the gelatin zymography as recently described [ ].

    Lysis:

    Article Title: Involvement of Akt/CREB signaling pathways in the protective effect of EPA against interleukin-1β-induced cytotoxicity and BDNF down-regulation in cultured rat hippocampal neurons
    Article Snippet: RIPA lysis buffer was purchased from Beyotime Biotechnology (Shanghai, China). .. Akt (catalog No. 9272), phospho-Akt (catalog No. 5012S, Ser473), CREB (catalog No. 4820) and phospho-CREB (catalog No. 9198, Ser133) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Article Title: MRP4 regulates ENaC-dependent CREB/COX-2/PGE2 signaling during embryo implantation
    Article Snippet: Immunoblotting The cells was lysed in ice-cold RIPA lysis buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.5% DOC, 0.1% SDS) with protease and phosphatase inhibitor cocktail (catalog #78443, Thermo Scientific) for 30 min on ice. .. Antibodies against MRP4 (1:100, Abcam, ab15602); phospho-CREB (1:1000, Cell Signaling, 9198), β-tubulin (1:2000, Santa Cruz, sc-9104) and β-actin (1:5000, Sigma, A1978).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Cell Signaling Technology Inc rabbit anti phospho creb
    The effect of N-methyl-D-aspartate receptor (NMDAR) blockade on the long-term plasticity, gene expression and protein synthesis. (A) In the presence of 2-amino-5-phosphonovaleric acid (APV), the plasticity map (that was obtained as in Figure 1 ) shows that TBS does no longer induce synaptic plasticity. The histogram confirms the almost complete absence of changes in the slice. (B) In the presence of APV, the pseudocolor maps of <t>P-CREB</t> and Creb levels (that were obtained as in Figure 2 ) do not show remarkable differences between a control slice and a slice that has received TBS. Histograms show that no significant differences occur between control slices and slices that have received TBS (mean and SEM, n = 4; 120 min). (C) In the presence of APV, the pseudocolor maps of <t>c-FOS</t> and c-Fos levels (that were obtained as in Figure 2 ) do not show remarkable differences between a control slice and a slice that has received TBS. Histograms show that no significant differences occur between control slices and slices that have received TBS (mean and SEM, n = 4; 120 min).
    Rabbit Anti Phospho Creb, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 85/100, based on 116 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti phospho creb/product/Cell Signaling Technology Inc
    Average 85 stars, based on 116 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho creb - by Bioz Stars, 2020-02
    85/100 stars
      Buy from Supplier

    90
    Cell Signaling Technology Inc rabbit mab phospho creb
    Effects of CSPGs on activities of p90RSK, <t>CREB</t> and LKB1 in N2A and CGN cultures. PTPσ/LAR transfected N2A cells or CGNs derived from PTPσ or LAR KO mice were treated with purified CSPGs (1.5 μg/ml) at several time points and the levels of phosphorylated p90RSK (p-p90RSK Thr573, active form), CREB (p-CREB <t>Ser133,</t> active form) and LKB1 (p-LKB1 Ser431, active form) in the supernatants of cell lysates were measured by Western blots. CSPGs did not alter levels of p-p90RSK in either PTPσ or LAR transfected N2A cells ( a ). CSPGs significantly decreased levels of p-CREB in PTPσ, not LAR, transfected N2A cells ( b ) and in PTPσ+/+, not PTPσ−/−, CGNs ( c ). In contrast, CSPGs stimulation decreased levels of p-LKB1 in LAR, not PTPσ, transfected N2A cells ( d ) and in LAR+/+, not LAR−/−, CGNs ( e ). The full-length blots are included in the Supplementary Information file.
    Rabbit Mab Phospho Creb, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit mab phospho creb/product/Cell Signaling Technology Inc
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rabbit mab phospho creb - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    Image Search Results


    The effect of N-methyl-D-aspartate receptor (NMDAR) blockade on the long-term plasticity, gene expression and protein synthesis. (A) In the presence of 2-amino-5-phosphonovaleric acid (APV), the plasticity map (that was obtained as in Figure 1 ) shows that TBS does no longer induce synaptic plasticity. The histogram confirms the almost complete absence of changes in the slice. (B) In the presence of APV, the pseudocolor maps of P-CREB and Creb levels (that were obtained as in Figure 2 ) do not show remarkable differences between a control slice and a slice that has received TBS. Histograms show that no significant differences occur between control slices and slices that have received TBS (mean and SEM, n = 4; 120 min). (C) In the presence of APV, the pseudocolor maps of c-FOS and c-Fos levels (that were obtained as in Figure 2 ) do not show remarkable differences between a control slice and a slice that has received TBS. Histograms show that no significant differences occur between control slices and slices that have received TBS (mean and SEM, n = 4; 120 min).

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Activation of the CREB/c-Fos Pathway during Long-Term Synaptic Plasticity in the Cerebellum Granular Layer

    doi: 10.3389/fncel.2017.00184

    Figure Lengend Snippet: The effect of N-methyl-D-aspartate receptor (NMDAR) blockade on the long-term plasticity, gene expression and protein synthesis. (A) In the presence of 2-amino-5-phosphonovaleric acid (APV), the plasticity map (that was obtained as in Figure 1 ) shows that TBS does no longer induce synaptic plasticity. The histogram confirms the almost complete absence of changes in the slice. (B) In the presence of APV, the pseudocolor maps of P-CREB and Creb levels (that were obtained as in Figure 2 ) do not show remarkable differences between a control slice and a slice that has received TBS. Histograms show that no significant differences occur between control slices and slices that have received TBS (mean and SEM, n = 4; 120 min). (C) In the presence of APV, the pseudocolor maps of c-FOS and c-Fos levels (that were obtained as in Figure 2 ) do not show remarkable differences between a control slice and a slice that has received TBS. Histograms show that no significant differences occur between control slices and slices that have received TBS (mean and SEM, n = 4; 120 min).

    Article Snippet: Immunohistochemistry Double immunofluorescent labeling was performed as follows: (1) sections were dried 30 min at room temperature and washed with Tris-buffered saline; (2) sections were blocked 1 h in Tris-buffered saline containing 10% normal horse serum (NHS) and 0.3% Triton X-100 (TX-100) at room temperature; (3) sections were incubated overnight at 4°C in Tris-buffered saline/1% NHS/0.3% TX-100 containing a mixture of a goat anti-c-FOS antibody (sc-52, diluted 1:400; Santa Cruz) and a rabbit anti-phospho-CREB (P-CREB, Ser133) antibody (87G3, diluted 1:100, Cell Signaling); (4) sections were rinsed in Tris-buffered saline and incubated 1 h at room temperature in Tris-buffered saline /1% NHS/0.3% TX-100 containing a mixture of Alexa Fluor 488 conjugated donkey anti-goat IgG antibody (1:300) and Alexa Fluor 594 conjugated donkey anti-rabbit IgG antibody (1:300, Life Technologies); and (5) sections were rinsed in Tris-buffered saline and covered with Prolong with DAPI.

    Techniques: Expressing

    PKA is not involved in GTN/H89-induced apoptosis (A) Immunoblot analysis of CREB and its phosphorylated form in SW480 cells exposed to 10 μM GTN and 10 μM H89 for 48 h. The immunoblot shown is representative of 3 independent experiments. (B) Exponentially growing SW480 cells (3 × 10 5 /mL) were treated with 10 μM GTN and/or 10 μM H89 or 1 μM KT5720 for 48 h at 37°C. Apoptotic cells were counted after Hoechst 33342 staining. (C) Cells were transfected with a PKA-specific siRNA (20 μM) for 6 h then treated with 10 μM GTN for 48 h at 37°C. Then apoptotic cells were counted. Results are the means of 3 independent experiments. Total protein was isolated and analyzed by immunoblot for expression of PKA-RIα, the active subunit of PKA, quantified relative to HSC70 expression. One immunoblot representative of 3 independent experiments is shown. (D) SW480 cells were treated with the following protein kinase inhibitors 1 h before exposure to 10 μM GTN: 20 μM SB203580, 15 μM PD98059, 1 nM rapamycin, 10 μM Y27632, and 10 μM H89. ‘All’ refers to treatment with all inhibitors except H89. Apoptotic cells were counted. Results are the means of 3 independent experiments.

    Journal: Oncotarget

    Article Title: H89 enhances the sensitivity of cancer cells to glyceryl trinitrate through a purinergic receptor-dependent pathway

    doi:

    Figure Lengend Snippet: PKA is not involved in GTN/H89-induced apoptosis (A) Immunoblot analysis of CREB and its phosphorylated form in SW480 cells exposed to 10 μM GTN and 10 μM H89 for 48 h. The immunoblot shown is representative of 3 independent experiments. (B) Exponentially growing SW480 cells (3 × 10 5 /mL) were treated with 10 μM GTN and/or 10 μM H89 or 1 μM KT5720 for 48 h at 37°C. Apoptotic cells were counted after Hoechst 33342 staining. (C) Cells were transfected with a PKA-specific siRNA (20 μM) for 6 h then treated with 10 μM GTN for 48 h at 37°C. Then apoptotic cells were counted. Results are the means of 3 independent experiments. Total protein was isolated and analyzed by immunoblot for expression of PKA-RIα, the active subunit of PKA, quantified relative to HSC70 expression. One immunoblot representative of 3 independent experiments is shown. (D) SW480 cells were treated with the following protein kinase inhibitors 1 h before exposure to 10 μM GTN: 20 μM SB203580, 15 μM PD98059, 1 nM rapamycin, 10 μM Y27632, and 10 μM H89. ‘All’ refers to treatment with all inhibitors except H89. Apoptotic cells were counted. Results are the means of 3 independent experiments.

    Article Snippet: After blocking non-specific binding sites for 2 h with 8% non fat milk in 0.1% Tween 20 in PBS (TPBS), the membrane was incubated overnight at 4°C with primary antibodies (Abs): anti-PARP (Cell Signaling, Saint Quentin Yvelines, France), anti-CREB (Euromedex, Mundolsheim, France), anti phospho-CREB (Cell Signaling), anti-PKA RIα (BD Transduction Laboratories, Le Pont de Claix, France) or anti-HSC-70 (Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Staining, Transfection, Isolation, Expressing

    Effects of CSPGs on activities of p90RSK, CREB and LKB1 in N2A and CGN cultures. PTPσ/LAR transfected N2A cells or CGNs derived from PTPσ or LAR KO mice were treated with purified CSPGs (1.5 μg/ml) at several time points and the levels of phosphorylated p90RSK (p-p90RSK Thr573, active form), CREB (p-CREB Ser133, active form) and LKB1 (p-LKB1 Ser431, active form) in the supernatants of cell lysates were measured by Western blots. CSPGs did not alter levels of p-p90RSK in either PTPσ or LAR transfected N2A cells ( a ). CSPGs significantly decreased levels of p-CREB in PTPσ, not LAR, transfected N2A cells ( b ) and in PTPσ+/+, not PTPσ−/−, CGNs ( c ). In contrast, CSPGs stimulation decreased levels of p-LKB1 in LAR, not PTPσ, transfected N2A cells ( d ) and in LAR+/+, not LAR−/−, CGNs ( e ). The full-length blots are included in the Supplementary Information file.

    Journal: Scientific Reports

    Article Title: Two PTP receptors mediate CSPG inhibition by convergent and divergent signaling pathways in neurons

    doi: 10.1038/srep37152

    Figure Lengend Snippet: Effects of CSPGs on activities of p90RSK, CREB and LKB1 in N2A and CGN cultures. PTPσ/LAR transfected N2A cells or CGNs derived from PTPσ or LAR KO mice were treated with purified CSPGs (1.5 μg/ml) at several time points and the levels of phosphorylated p90RSK (p-p90RSK Thr573, active form), CREB (p-CREB Ser133, active form) and LKB1 (p-LKB1 Ser431, active form) in the supernatants of cell lysates were measured by Western blots. CSPGs did not alter levels of p-p90RSK in either PTPσ or LAR transfected N2A cells ( a ). CSPGs significantly decreased levels of p-CREB in PTPσ, not LAR, transfected N2A cells ( b ) and in PTPσ+/+, not PTPσ−/−, CGNs ( c ). In contrast, CSPGs stimulation decreased levels of p-LKB1 in LAR, not PTPσ, transfected N2A cells ( d ) and in LAR+/+, not LAR−/−, CGNs ( e ). The full-length blots are included in the Supplementary Information file.

    Article Snippet: Sources of compounds Antibodies against the following proteins were used: mouse mAb phospho-Akt (Ser473, 587F11), rabbit mAb phospho-S6 ribosomal protein (Ser235/236), rabbit mAb phospho-Erk1/2 (p44/42), rabbit pAb phospho-CRMP2 (Thr514), rabbit pAb phospho-cofilin (Ser3), rabbit mAb phospho-4E- BP1 (eukaryotic initiation factor 4E-binding protein 1, Thr37/46, 236B4), rabbit mAb phospho-CREB (Ser133, 87G3), rabbit mAb phospho-PKA C (Thr197, D45D3), rabbit pAb phospho-90 kDa ribosomal S6 kinase (p90RSK, Thr573), rabbit pAb phospho-PKCζ/λ (Thr410/403), rabbit mAb phospho-PKC (pan) (zeta Thr410, 190D10), rabbit mAb phospho-LKB1 (Ser428, C67A3) (all from Cell Signaling Technology), rabbit pAb phospho-MAP1B (Thr1265, Millipore), rabbit pAb phospho-APC (Ser2054, Abcam), mouse anti-actin clone C4 (MP Biomedicals) and mouse mAb against RhoA (sc-418; from Santa Cruz Biotechnology).

    Techniques: Transfection, Derivative Assay, Mouse Assay, Purification, Western Blot

    Expression and activation state of signalling mediators involved in regulation of smooth muscle tone. Western blot analysis was performed for cortical and subcortical brain arterioles from 13 sheep, as described in the methods section. Differences of expression levels of signalling proteins involved in control of smooth muscle tone: (A) nNOS, P-eNOS and eNOS-protein, (B) P-CREB and CREB protein, (C) P-ERK and ERK protein was detected in relation to β-actin. As these data were not normally distributed they are presented as box plots, where boxes represent 25th and 75th percentiles, respectively. Medians are indicated by horizontal lines. Whiskers indicate 10th and 90th percentiles, respectively. 1+2, samples from two different sheep; Cx, cortex; Scx, subcortex; AU, arbitrary units; Ref. reference sample.

    Journal: PLoS ONE

    Article Title: Underlying mechanism of subcortical brain protection during hypoxia and reoxygenation in a sheep model - Influence of α1-adrenergic signalling

    doi: 10.1371/journal.pone.0196363

    Figure Lengend Snippet: Expression and activation state of signalling mediators involved in regulation of smooth muscle tone. Western blot analysis was performed for cortical and subcortical brain arterioles from 13 sheep, as described in the methods section. Differences of expression levels of signalling proteins involved in control of smooth muscle tone: (A) nNOS, P-eNOS and eNOS-protein, (B) P-CREB and CREB protein, (C) P-ERK and ERK protein was detected in relation to β-actin. As these data were not normally distributed they are presented as box plots, where boxes represent 25th and 75th percentiles, respectively. Medians are indicated by horizontal lines. Whiskers indicate 10th and 90th percentiles, respectively. 1+2, samples from two different sheep; Cx, cortex; Scx, subcortex; AU, arbitrary units; Ref. reference sample.

    Article Snippet: These include: anti-NOS1 (nNOS; sc-8309 (H-299)) and anti-NOS3 (eNOS; sc-8311 (H-159)), both from Santa Cruz Biotechnology (Dallas, TX, USA); anti-phospho-eNOS (P-Ser1177; 9570), anti-CREB (rabbit monoclonal 48H2; 9197), anti-phospho-CREB (P-Ser133; rabbit monoclonal 87G3; 9198), anti-ERK1/2 (rabbit monoclonal 137F5; 4695) and anti-phospho-ERK1/2 (P-Thr202/Tyr204; rabbit monoclonal D13.14.4; 4370), all from Cell Signaling Technologies (Frankfurt am Main, Germany).

    Techniques: Expressing, Activation Assay, Western Blot