electrospray ionization tandem mass spectrometry individual chemical standards  (Millipore)


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    Name:
    Mass Spectrometry
    Description:
    This reference summarizes the theory instrumentation and application of mass spectrometry and atmospheric pressure ionization to screen evaluate and improve the performance and quality of drug candidates It covers recent advances in combinatorial chemistry molecular biology bioanalysis automation and computing cassette dosing the utilization of liquid chromatography mass spectrometry LC MS in chemical library characterization combinatorial chemistry drug transport and in vivo microdialysis reliable methods of sample preparation and handling novel strategies to determine and characterize metabolite structures the impact of absorption distribution metabolism and elimination properties on drug discovery
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    Millipore electrospray ionization tandem mass spectrometry individual chemical standards
    Mass Spectrometry
    This reference summarizes the theory instrumentation and application of mass spectrometry and atmospheric pressure ionization to screen evaluate and improve the performance and quality of drug candidates It covers recent advances in combinatorial chemistry molecular biology bioanalysis automation and computing cassette dosing the utilization of liquid chromatography mass spectrometry LC MS in chemical library characterization combinatorial chemistry drug transport and in vivo microdialysis reliable methods of sample preparation and handling novel strategies to determine and characterize metabolite structures the impact of absorption distribution metabolism and elimination properties on drug discovery
    https://www.bioz.com/result/electrospray ionization tandem mass spectrometry individual chemical standards/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    electrospray ionization tandem mass spectrometry individual chemical standards - by Bioz Stars, 2020-09
    99/100 stars

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    Related Articles

    Transfection:

    Article Title: Cdk2 and Pin1 negatively regulate the transcriptional corepressor SMRT
    Article Snippet: .. Mass spectrometry HeLa WCEs transfected with FLAG-SMRT (1,178–1,823) and treated with 20 nM calyculin A were subjected to coimmunoprecipitations using anti-FLAG affinity beads (Sigma-Aldrich) and separated by SDS-PAGE. .. The gel was Coomassie stained, and protein bands were cut out, washed and destained (50% ethanol and 5% acetic acid), reduced and alkylated (DTT and iodoacetamide), dehydrated (acetonitrile), and dried in a speed vac.

    Article Title: 14–3-3ζ inhibits heme oxygenase-1 (HO-1) degradation and promotes hepatocellular carcinoma proliferation: involvement of STAT3 signaling
    Article Snippet: .. Mass spectrometry (MS) Whole cell lysates from 293FT cells transient transfected with FLAG-HO-1 or FLAG-vector were lysed in lysis buffer (50 mM Tris-HCl (pH 7.4), 100 mM NaCl, 0.5% NP-40, 1 mM EDTA and protease inhibitor cocktails) and applied to an IP lysis buffer equilibrated FLAG affinity gel (Sigma-Aldrich, St. Louis, MO, USA) in 4 °C for 4 h. After binding, the affinity gels were washed with the lysis buffer for five times and the pellets were resuspended in SDS sample buffer and boiled for 8 min. ..

    Silver Staining:

    Article Title: Hyperphosphatemia, Phosphoprotein Phosphatases, and Microparticle Release in Vascular Endothelial Cells
    Article Snippet: .. The 2-DE gels were stained by Silver Stain Plus (Bio-Rad) or RAPIDstain Reagent (Calbiochem; if 2-DE was to be followed by mass spectrometry). .. In some experiments, proteins from 2-DE gels were blotted onto nitrocellulose membranes (Amersham) followed by probing with primary pan-specific antibody against P-Ser/Thr (Antibodies-Online; Beckton Dickinson).

    Sample Prep:

    Article Title: HMGB1 Release and Redox Regulates Autophagy and Apoptosis in Cancer Cells
    Article Snippet: .. MALDI-TOF mass spectrometry For MALDI-TOF Mass Spectrometry sample preparation, solid-phase extraction pipette tips (C18 ZipTip) from Millipore were used. .. Protein sequencing grade trifluoroacetic acid (TFA) was purchased from Fisher Scientific.

    Protease Inhibitor:

    Article Title: 14–3-3ζ inhibits heme oxygenase-1 (HO-1) degradation and promotes hepatocellular carcinoma proliferation: involvement of STAT3 signaling
    Article Snippet: .. Mass spectrometry (MS) Whole cell lysates from 293FT cells transient transfected with FLAG-HO-1 or FLAG-vector were lysed in lysis buffer (50 mM Tris-HCl (pH 7.4), 100 mM NaCl, 0.5% NP-40, 1 mM EDTA and protease inhibitor cocktails) and applied to an IP lysis buffer equilibrated FLAG affinity gel (Sigma-Aldrich, St. Louis, MO, USA) in 4 °C for 4 h. After binding, the affinity gels were washed with the lysis buffer for five times and the pellets were resuspended in SDS sample buffer and boiled for 8 min. ..

    Transferring:

    Article Title: HMGB1 Release and Redox Regulates Autophagy and Apoptosis in Cancer Cells
    Article Snippet: .. MALDI-TOF mass spectrometry For MALDI-TOF Mass Spectrometry sample preparation, solid-phase extraction pipette tips (C18 ZipTip) from Millipore were used. .. Protein sequencing grade trifluoroacetic acid (TFA) was purchased from Fisher Scientific.

    Purification:

    Article Title: Decreased levels of genuine large free hCG alpha in men presenting with abnormal semen analysis
    Article Snippet: .. Verification of hCGα by mass spectrometry hCGα purified from seminal plasma by IP, as described above (2.4.), was analyzed by SDS-PAGE, and the protein band at 24 kDa was excised from the gel and digested with endoproteinase Lys-C [EC 3.4.21.50] (Sigma-Aldrich, 1/20 w/w) in 100 mM H4 HCO3 buffer (pH = 8.0) for 2 hours at 37°C. .. The digest was analyzed using nano-HPLC consisting of an UltiMate 3000 System (Dionex Corporation) connected online to a linear iontrap mass spectrometer ThermoElectron Finnigan LTQ) equipped with a nanospray ionization source.

    Mass Spectrometry:

    Article Title: HMGB1 Release and Redox Regulates Autophagy and Apoptosis in Cancer Cells
    Article Snippet: .. MALDI-TOF mass spectrometry For MALDI-TOF Mass Spectrometry sample preparation, solid-phase extraction pipette tips (C18 ZipTip) from Millipore were used. .. Protein sequencing grade trifluoroacetic acid (TFA) was purchased from Fisher Scientific.

    Article Title: Selective Inhibition on Organic Cation Transporters by Carvedilol Protects Mice from Cisplatin-Induced Nephrotoxicity
    Article Snippet: .. The nitric acid used to lysate cells and the chemical standards for cisplatin measurement by inductively coupled plasma mass spectrometry (ICP-MS) were of trace ICP-MS grade and supplied by Sigma-Aldrich Corp. (St Louis, MO). ..

    Article Title: Decreased levels of genuine large free hCG alpha in men presenting with abnormal semen analysis
    Article Snippet: .. Verification of hCGα by mass spectrometry hCGα purified from seminal plasma by IP, as described above (2.4.), was analyzed by SDS-PAGE, and the protein band at 24 kDa was excised from the gel and digested with endoproteinase Lys-C [EC 3.4.21.50] (Sigma-Aldrich, 1/20 w/w) in 100 mM H4 HCO3 buffer (pH = 8.0) for 2 hours at 37°C. .. The digest was analyzed using nano-HPLC consisting of an UltiMate 3000 System (Dionex Corporation) connected online to a linear iontrap mass spectrometer ThermoElectron Finnigan LTQ) equipped with a nanospray ionization source.

    Article Title: Hyperphosphatemia, Phosphoprotein Phosphatases, and Microparticle Release in Vascular Endothelial Cells
    Article Snippet: .. The 2-DE gels were stained by Silver Stain Plus (Bio-Rad) or RAPIDstain Reagent (Calbiochem; if 2-DE was to be followed by mass spectrometry). .. In some experiments, proteins from 2-DE gels were blotted onto nitrocellulose membranes (Amersham) followed by probing with primary pan-specific antibody against P-Ser/Thr (Antibodies-Online; Beckton Dickinson).

    Article Title: Cdk2 and Pin1 negatively regulate the transcriptional corepressor SMRT
    Article Snippet: .. Mass spectrometry HeLa WCEs transfected with FLAG-SMRT (1,178–1,823) and treated with 20 nM calyculin A were subjected to coimmunoprecipitations using anti-FLAG affinity beads (Sigma-Aldrich) and separated by SDS-PAGE. .. The gel was Coomassie stained, and protein bands were cut out, washed and destained (50% ethanol and 5% acetic acid), reduced and alkylated (DTT and iodoacetamide), dehydrated (acetonitrile), and dried in a speed vac.

    Article Title: Molecular basis for prey relocation in viperid snakes
    Article Snippet: .. Mass determination by mass spectrometry (MALDI-TOF-TOF) Peak III from size exclusion (BioGel P-100 column) was desalted using C4 ZipTips (Millipore Inc., Billerica, MA, USA) and analyzed using a Bruker Ultraflex MALDI-TOF mass spectrometer (Proteomics and Metabolomics Facility, Colorado State University, Fort Collins, CO, USA) operating in linear mode. ..

    Article Title: 14–3-3ζ inhibits heme oxygenase-1 (HO-1) degradation and promotes hepatocellular carcinoma proliferation: involvement of STAT3 signaling
    Article Snippet: .. Mass spectrometry (MS) Whole cell lysates from 293FT cells transient transfected with FLAG-HO-1 or FLAG-vector were lysed in lysis buffer (50 mM Tris-HCl (pH 7.4), 100 mM NaCl, 0.5% NP-40, 1 mM EDTA and protease inhibitor cocktails) and applied to an IP lysis buffer equilibrated FLAG affinity gel (Sigma-Aldrich, St. Louis, MO, USA) in 4 °C for 4 h. After binding, the affinity gels were washed with the lysis buffer for five times and the pellets were resuspended in SDS sample buffer and boiled for 8 min. ..

    Article Title: Dihydrofolate Reductase/Thymidylate Synthase Fine-Tunes the Folate Status and Controls Redox Homeostasis in Plants
    Article Snippet: .. For long-term treatments, seeds were plated on control MS/2 media or on MS/2 media supplemented with 30 µM TMP (Trimethoprim; Sigma-Aldrich), 1 mM HU (Sigma-Aldrich), or both. .. For short-term treatments, 3-d-old seedlings were cultured in liquid MS/2 media supplemented with 5 mM HU for 5 h.

    Staining:

    Article Title: Hyperphosphatemia, Phosphoprotein Phosphatases, and Microparticle Release in Vascular Endothelial Cells
    Article Snippet: .. The 2-DE gels were stained by Silver Stain Plus (Bio-Rad) or RAPIDstain Reagent (Calbiochem; if 2-DE was to be followed by mass spectrometry). .. In some experiments, proteins from 2-DE gels were blotted onto nitrocellulose membranes (Amersham) followed by probing with primary pan-specific antibody against P-Ser/Thr (Antibodies-Online; Beckton Dickinson).

    Lysis:

    Article Title: 14–3-3ζ inhibits heme oxygenase-1 (HO-1) degradation and promotes hepatocellular carcinoma proliferation: involvement of STAT3 signaling
    Article Snippet: .. Mass spectrometry (MS) Whole cell lysates from 293FT cells transient transfected with FLAG-HO-1 or FLAG-vector were lysed in lysis buffer (50 mM Tris-HCl (pH 7.4), 100 mM NaCl, 0.5% NP-40, 1 mM EDTA and protease inhibitor cocktails) and applied to an IP lysis buffer equilibrated FLAG affinity gel (Sigma-Aldrich, St. Louis, MO, USA) in 4 °C for 4 h. After binding, the affinity gels were washed with the lysis buffer for five times and the pellets were resuspended in SDS sample buffer and boiled for 8 min. ..

    Binding Assay:

    Article Title: 14–3-3ζ inhibits heme oxygenase-1 (HO-1) degradation and promotes hepatocellular carcinoma proliferation: involvement of STAT3 signaling
    Article Snippet: .. Mass spectrometry (MS) Whole cell lysates from 293FT cells transient transfected with FLAG-HO-1 or FLAG-vector were lysed in lysis buffer (50 mM Tris-HCl (pH 7.4), 100 mM NaCl, 0.5% NP-40, 1 mM EDTA and protease inhibitor cocktails) and applied to an IP lysis buffer equilibrated FLAG affinity gel (Sigma-Aldrich, St. Louis, MO, USA) in 4 °C for 4 h. After binding, the affinity gels were washed with the lysis buffer for five times and the pellets were resuspended in SDS sample buffer and boiled for 8 min. ..

    SDS Page:

    Article Title: Decreased levels of genuine large free hCG alpha in men presenting with abnormal semen analysis
    Article Snippet: .. Verification of hCGα by mass spectrometry hCGα purified from seminal plasma by IP, as described above (2.4.), was analyzed by SDS-PAGE, and the protein band at 24 kDa was excised from the gel and digested with endoproteinase Lys-C [EC 3.4.21.50] (Sigma-Aldrich, 1/20 w/w) in 100 mM H4 HCO3 buffer (pH = 8.0) for 2 hours at 37°C. .. The digest was analyzed using nano-HPLC consisting of an UltiMate 3000 System (Dionex Corporation) connected online to a linear iontrap mass spectrometer ThermoElectron Finnigan LTQ) equipped with a nanospray ionization source.

    Article Title: Cdk2 and Pin1 negatively regulate the transcriptional corepressor SMRT
    Article Snippet: .. Mass spectrometry HeLa WCEs transfected with FLAG-SMRT (1,178–1,823) and treated with 20 nM calyculin A were subjected to coimmunoprecipitations using anti-FLAG affinity beads (Sigma-Aldrich) and separated by SDS-PAGE. .. The gel was Coomassie stained, and protein bands were cut out, washed and destained (50% ethanol and 5% acetic acid), reduced and alkylated (DTT and iodoacetamide), dehydrated (acetonitrile), and dried in a speed vac.

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  • 99
    Millipore tmz
    Therapeutic effect of <t>ALZ003</t> on the growth of <t>TMZ-resistant</t> glioblastoma in vivo . A. After intracranial transplantation with TMZ-resistant U87MG cells for 10 days, mice were orally and intravenously administrated with TMZ (15 mg/kg) and ALZ003 three times per week, respectively. The date of death was recorded and analyzed by Log-Rank Test. (** p = 0.0052). B. The mouse brains were paraffin embedded and subject to slides preparation followed by hematoxylin- and eosin-staining. Tumor in each slide was indicated. The scale bar for HE staining of whole brain slice is 1 mm, and that for other IHC images is 0.5 mm. C. The area of tumor was quantitated by the Image J software. The difference with the control group was analyzed using Student's t -test (* p
    Tmz, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tmz/product/Millipore
    Average 99 stars, based on 70 article reviews
    Price from $9.99 to $1999.99
    tmz - by Bioz Stars, 2020-09
    99/100 stars
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    99
    Millipore pkg inhibitor
    Schematic representation of <t>BNP</t> signaling pathway leading to Sca-1 + cell proliferation. ( A ) Number of sorted Sca-1 high+ cells after 11 days of culture with or without BNP or CNP and <t>PKG</t> or p38 inhibitor. n = at least 6 different experiments per group. The data were related to the average of untreated cells. All the results are means ± SEM, *p
    Pkg Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pkg inhibitor/product/Millipore
    Average 99 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
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    95
    Millipore esi 09
    Effect of <t>ESI-09</t> and HJC0197, EPAC inhibitors, and KT-5720 and PKI, a PKA inhibitors, on PAK4 activation ( A ), CREB phosphorylation ( B ), p44/42 phosphorylation ( C ), and p38 phosphorylation ( D ) by VIP in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of ESI-09 (100 µM), HJC0197 (10 µM, 2 h), KT-5720 (10 µM) or PKI (10 µM) for 1 h and then incubated with no addition (control) or VIP (10 nM) for 15 min, and then lysed. Western blots were analyzed using anti-pS474 PAK4, pS133 CREB, pT202/Y204 p44/42 or pT180/Y182 p38 and, as loading control, antitotal PAK4, antitotal CREB, antitotal p44/42, and antitotal p38. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of basal stimulation of the control group. * P
    Esi 09, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    esi 09 - by Bioz Stars, 2020-09
    95/100 stars
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    Image Search Results


    Therapeutic effect of ALZ003 on the growth of TMZ-resistant glioblastoma in vivo . A. After intracranial transplantation with TMZ-resistant U87MG cells for 10 days, mice were orally and intravenously administrated with TMZ (15 mg/kg) and ALZ003 three times per week, respectively. The date of death was recorded and analyzed by Log-Rank Test. (** p = 0.0052). B. The mouse brains were paraffin embedded and subject to slides preparation followed by hematoxylin- and eosin-staining. Tumor in each slide was indicated. The scale bar for HE staining of whole brain slice is 1 mm, and that for other IHC images is 0.5 mm. C. The area of tumor was quantitated by the Image J software. The difference with the control group was analyzed using Student's t -test (* p

    Journal: Redox Biology

    Article Title: AR ubiquitination induced by the curcumin analog suppresses growth of temozolomide-resistant glioblastoma through disrupting GPX4-Mediated redox homeostasis

    doi: 10.1016/j.redox.2019.101413

    Figure Lengend Snippet: Therapeutic effect of ALZ003 on the growth of TMZ-resistant glioblastoma in vivo . A. After intracranial transplantation with TMZ-resistant U87MG cells for 10 days, mice were orally and intravenously administrated with TMZ (15 mg/kg) and ALZ003 three times per week, respectively. The date of death was recorded and analyzed by Log-Rank Test. (** p = 0.0052). B. The mouse brains were paraffin embedded and subject to slides preparation followed by hematoxylin- and eosin-staining. Tumor in each slide was indicated. The scale bar for HE staining of whole brain slice is 1 mm, and that for other IHC images is 0.5 mm. C. The area of tumor was quantitated by the Image J software. The difference with the control group was analyzed using Student's t -test (* p

    Article Snippet: Both of TMZ and ALZ003 were dissolved in dimethyl sulfoxide (DMSO, MilliporeSigma Corporate), and stored at −20 °C. siRNA targeting AR or GPX4 was purchased from GE Healthcare Dharmacon, Inc. (Lafayette, CO, USA).

    Techniques: In Vivo, Transplantation Assay, Mouse Assay, Staining, Slice Preparation, Immunohistochemistry, Software

    Schematic representation of BNP signaling pathway leading to Sca-1 + cell proliferation. ( A ) Number of sorted Sca-1 high+ cells after 11 days of culture with or without BNP or CNP and PKG or p38 inhibitor. n = at least 6 different experiments per group. The data were related to the average of untreated cells. All the results are means ± SEM, *p

    Journal: Scientific Reports

    Article Title: Natriuretic Peptide Receptor B modulates the proliferation of the cardiac cells expressing the Stem Cell Antigen-1

    doi: 10.1038/srep41936

    Figure Lengend Snippet: Schematic representation of BNP signaling pathway leading to Sca-1 + cell proliferation. ( A ) Number of sorted Sca-1 high+ cells after 11 days of culture with or without BNP or CNP and PKG or p38 inhibitor. n = at least 6 different experiments per group. The data were related to the average of untreated cells. All the results are means ± SEM, *p

    Article Snippet: To determine by which signaling pathway BNP acts on Sca-1high+ cells to induce their proliferation, a PKG inhibitor (Guanosine 3′, 5′-cyclic Monophosphorothioate, β-Phenyl-1, N2-etheno-8-bromo-Rp-Isomer, Sodium Salt (Calbiochem) 0.1 microM)) or a p38 MAP kinase inhibitor (SB203580 (Enzo) 10 microM) was added to the cell culture 15 min or 1 hour, respectively, before the BNP treatment.

    Techniques:

    RASIP1 is a RAP1 effector that controls EC junction morphology. (A) Pull-down assay using purified RAP1A, RHOA, and RAC1 GST fusion proteins with control (c) or RASIP1 shRNA (kd) HUVEC lysates. RASIP1 interacts weakly with RAP1A-GST loaded with GDP, and

    Journal: Blood

    Article Title: Rasip1 regulates vertebrate vascular endothelial junction stability through Epac1-Rap1 signaling

    doi: 10.1182/blood-2013-02-483156

    Figure Lengend Snippet: RASIP1 is a RAP1 effector that controls EC junction morphology. (A) Pull-down assay using purified RAP1A, RHOA, and RAC1 GST fusion proteins with control (c) or RASIP1 shRNA (kd) HUVEC lysates. RASIP1 interacts weakly with RAP1A-GST loaded with GDP, and

    Article Snippet: Purified RAP1A–glutathione S-transferase (GST) (Sigma-Aldrich), Ras homolog gene family member A (RHOA)-GST, and RAC1A-GST (Novus) were incubated in cell lysis buffer with 10 μM guanosine diphosphate (GDP) or GTPγS (Millipore) for 30 minutes at 37°C.

    Techniques: Pull Down Assay, Purification, shRNA

    Effect of ESI-09 and HJC0197, EPAC inhibitors, and KT-5720 and PKI, a PKA inhibitors, on PAK4 activation ( A ), CREB phosphorylation ( B ), p44/42 phosphorylation ( C ), and p38 phosphorylation ( D ) by VIP in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of ESI-09 (100 µM), HJC0197 (10 µM, 2 h), KT-5720 (10 µM) or PKI (10 µM) for 1 h and then incubated with no addition (control) or VIP (10 nM) for 15 min, and then lysed. Western blots were analyzed using anti-pS474 PAK4, pS133 CREB, pT202/Y204 p44/42 or pT180/Y182 p38 and, as loading control, antitotal PAK4, antitotal CREB, antitotal p44/42, and antitotal p38. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of basal stimulation of the control group. * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Hormones, Neurotransmitters, Growth Factors, Receptors, and Signaling: Cyclic AMP-dependent protein kinase A and EPAC mediate VIP and secretin stimulation of PAK4 and activation of Na+,K+-ATPase in pancreatic acinar cells

    doi: 10.1152/ajpgi.00275.2018

    Figure Lengend Snippet: Effect of ESI-09 and HJC0197, EPAC inhibitors, and KT-5720 and PKI, a PKA inhibitors, on PAK4 activation ( A ), CREB phosphorylation ( B ), p44/42 phosphorylation ( C ), and p38 phosphorylation ( D ) by VIP in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of ESI-09 (100 µM), HJC0197 (10 µM, 2 h), KT-5720 (10 µM) or PKI (10 µM) for 1 h and then incubated with no addition (control) or VIP (10 nM) for 15 min, and then lysed. Western blots were analyzed using anti-pS474 PAK4, pS133 CREB, pT202/Y204 p44/42 or pT180/Y182 p38 and, as loading control, antitotal PAK4, antitotal CREB, antitotal p44/42, and antitotal p38. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of basal stimulation of the control group. * P

    Article Snippet: ESI-09 and protein kinase A (PKA) inhibitor 14–22 Amide PKA Type II inhibitor (PKI) were from Millipore (Billerica, MA).

    Techniques: Activation Assay, Isolation, Incubation, Western Blot

    Effect of ESI-09 and HJC0197, EPAC inhibitors, and KT-5720 and PKI, PKA inhibitors, on PAK4 activation ( A ), CREB phosphorylation ( B ), p44/42 phosphorylation ( C ), and p38 phosphorylation ( D ) by secretin in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of ESI-09 (100 µM), HJC0197 (10 µM, 2h), KT-5720 (10 µM), or PKI (10 µM) for 1 h and then incubated with no addition (control) or secretin (10 nM) for 15 min, and then lysed. Western blots were analyzed using anti-pS474 PAK4, pS133 CREB, pT202/Y204 p44/42, or pT180/Y182 p38 and, as loading control, antitotal PAK4, antitotal CREB, antitotal p44/42, and antitotal p38. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of basal stimulation of the control group. * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Hormones, Neurotransmitters, Growth Factors, Receptors, and Signaling: Cyclic AMP-dependent protein kinase A and EPAC mediate VIP and secretin stimulation of PAK4 and activation of Na+,K+-ATPase in pancreatic acinar cells

    doi: 10.1152/ajpgi.00275.2018

    Figure Lengend Snippet: Effect of ESI-09 and HJC0197, EPAC inhibitors, and KT-5720 and PKI, PKA inhibitors, on PAK4 activation ( A ), CREB phosphorylation ( B ), p44/42 phosphorylation ( C ), and p38 phosphorylation ( D ) by secretin in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of ESI-09 (100 µM), HJC0197 (10 µM, 2h), KT-5720 (10 µM), or PKI (10 µM) for 1 h and then incubated with no addition (control) or secretin (10 nM) for 15 min, and then lysed. Western blots were analyzed using anti-pS474 PAK4, pS133 CREB, pT202/Y204 p44/42, or pT180/Y182 p38 and, as loading control, antitotal PAK4, antitotal CREB, antitotal p44/42, and antitotal p38. Bands were visualized using chemiluminescence and quantified by densitometry. Top : results of a representative blot of 3 independent experiments are shown. Bottom : means ± SE of at least 4 independent experiments. Results are expressed as percentages of basal stimulation of the control group. * P

    Article Snippet: ESI-09 and protein kinase A (PKA) inhibitor 14–22 Amide PKA Type II inhibitor (PKI) were from Millipore (Billerica, MA).

    Techniques: Activation Assay, Isolation, Incubation, Western Blot