P. falciparum merozoites were isolated in IC buffer and treated for 15 mins with the following at concentrations shown: 100 µM IBMX (3-isobutyl-1-methylxanthine, Calbiochem, USA), 50 µM KH7 (Calbiochem, USA), 100 µM ACTZ (acetazolamide, Sigma, USA), 20 µM DiB-cAMP (adenosine 3,5 cyclic monophosphate, N6, O2- dibutyryl sodium salt, Calbiochem, USA), 10 µM Ca2+ ionophore A23187 (Calbiochem, USA), 50 µM BAPTA-AM (Calbiochem, USA), 10 µM U73122 (Calbiochem, USA), 10 µM U73343 (Calbiochem, USA), 200 µM Epac agonist (8-pCPT-2’-O-Me-cAMP, Sigma, USA), 25 µM Epac antagonist ESI-05 (Biolog, Germany), 25 µM Epac antagonist ESI-09 (Biolog, Germany) and 50 µM geranyl geranyl transferase inhibitor, GGTI 298, which disrupts Rap1 function. Merozoites were then transferred to EC buffer (5 mM KCl, 140 mM NaCl, 1 mM MgCl2, 2 mM EGTA, 5.6 mM glucose, 25 mM HEPES, pH 7.2) followed by either measurement of cytosolic cAMP and Ca2+ levels in merozoites or microneme secretion.
Esi 05
Manufactured by Biolog
Sourced in Germany, United States
ESI-05 is an electrochemical sensor that measures the electrical properties of a sample. It is designed to provide accurate and reliable measurements of various electrochemical parameters, such as conductivity, pH, and redox potential. The ESI-05 is a versatile piece of lab equipment suitable for a wide range of applications in various industries.
Automatically generated - may contain errors
Lab products found in correlation
Esi 09, by Biolog (2 mentions)
Dexrazoxane, by Merck Group (2 mentions)
Zvad fmk, by Bachem (2 mentions)
8 pcpt 2 o me camp, by Merck Group (1 mentions)
Anti phospho creb ser133, by Cell Signaling Technology (1 mentions)
Anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Peroxidase conjugated anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Anti rap1, by Merck Group (1 mentions)
Alexa fluor 488 conjugated anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
Anti creb, by Cell Signaling Technology (1 mentions)
Glucagon, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Ggti 298, by Bio-Techne (1 mentions)
Thapsigargin, by Bio-Techne (1 mentions)
Isradipine, by Alomone (1 mentions)
Bafilomycin, by Bio-Techne (1 mentions)
Noradrenaline, by Bio-Techne (1 mentions)
Prazosin, by Abcam (1 mentions)
Ned 19, by Bio-Techne (1 mentions)
Xestospongin c, by Bio-Techne (1 mentions)
10 protocols using esi 05
1
Modulation of Malaria Merozoite Signaling
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P. falciparum merozoites were isolated in IC buffer and treated for 15 mins with the following at concentrations shown: 100 µM IBMX (3-isobutyl-1-methylxanthine, Calbiochem, USA), 50 µM KH7 (Calbiochem, USA), 100 µM ACTZ (acetazolamide, Sigma, USA), 20 µM DiB-cAMP (adenosine 3,5 cyclic monophosphate, N6, O2- dibutyryl sodium salt, Calbiochem, USA), 10 µM Ca2+ ionophore A23187 (Calbiochem, USA), 50 µM BAPTA-AM (Calbiochem, USA), 10 µM U73122 (Calbiochem, USA), 10 µM U73343 (Calbiochem, USA), 200 µM Epac agonist (8-pCPT-2’-O-Me-cAMP, Sigma, USA), 25 µM Epac antagonist ESI-05 (Biolog, Germany), 25 µM Epac antagonist ESI-09 (Biolog, Germany) and 50 µM geranyl geranyl transferase inhibitor, GGTI 298, which disrupts Rap1 function. Merozoites were then transferred to EC buffer (5 mM KCl, 140 mM NaCl, 1 mM MgCl2, 2 mM EGTA, 5.6 mM glucose, 25 mM HEPES, pH 7.2) followed by either measurement of cytosolic cAMP and Ca2+ levels in merozoites or microneme secretion.
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P. falciparum merozoites were isolated in IC buffer and treated for 15 mins with the following at concentrations shown: 100 µM IBMX (3-isobutyl-1-methylxanthine, Calbiochem, USA), 50 µM KH7 (Calbiochem, USA), 100 µM ACTZ (acetazolamide, Sigma, USA), 20 µM DiB-cAMP (adenosine 3,5 cyclic monophosphate, N6, O2- dibutyryl sodium salt, Calbiochem, USA), 10 µM Ca2+ ionophore A23187 (Calbiochem, USA), 50 µM BAPTA-AM (Calbiochem, USA), 10 µM U73122 (Calbiochem, USA), 10 µM U73343 (Calbiochem, USA), 200 µM Epac agonist (8-pCPT-2’-O-Me-cAMP, Sigma, USA), 25 µM Epac antagonist ESI-05 (Biolog, Germany), 25 µM Epac antagonist ESI-09 (Biolog, Germany) and 50 µM geranyl geranyl transferase inhibitor, GGTI 298, which disrupts Rap1 function. Merozoites were then transferred to EC buffer (5 mM KCl, 140 mM NaCl, 1 mM MgCl2, 2 mM EGTA, 5.6 mM glucose, 25 mM HEPES, pH 7.2) followed by either measurement of cytosolic cAMP and Ca2+ levels in merozoites or microneme secretion.
2
Measuring P. falciparum Merozoite Invasion Rates
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P. falciparum merozoites isolated as described above were either mock-treated or treated with 50 µm KH7 (Calbiochem, USA), 100 µm ACTZ (Sigma, USA), 25 µM ESI-05 (Biolog, Germany) or 25 µM ESI-09 (Biolog, Germany) for 15 min at 37°C, washed with IC buffer, resuspended in EC buffer and incubated with erythrocytes in EC buffer at 37°C under mixed gas environment for 2 h to allow invasion. EC buffer was then replaced with complete RPMI. After 18–20 h of incubation in complete RPMI under mixed gas environment to allow development of ring stages, the percentage of infected erythrocytes was scored by flow cytometry to determine invasion rates, as described earlier [52] (link).
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P. falciparum merozoites isolated as described above were either mock-treated or treated with 50 µm KH7 (Calbiochem, USA), 100 µm ACTZ (Sigma, USA), 25 µM ESI-05 (Biolog, Germany) or 25 µM ESI-09 (Biolog, Germany) for 15 min at 37°C, washed with IC buffer, resuspended in EC buffer and incubated with erythrocytes in EC buffer at 37°C under mixed gas environment for 2 h to allow invasion. EC buffer was then replaced with complete RPMI. After 18–20 h of incubation in complete RPMI under mixed gas environment to allow development of ring stages, the percentage of infected erythrocytes was scored by flow cytometry to determine invasion rates, as described earlier [52] (link).
3
Signaling Pathway Analysis Protocol
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Glucagon and insulin were purchased from Sigma‐Aldrich (St Louis, MO, USA); 8‐CPT‐2‐O‐Me‐cAMP (8‐CPT), 6‐Bnz‐cAMP, and ESI‐05 were purchased from Biolog (Bremen, Germany). GP inhibitor (GPi) was purchased from Calbiochem (San Diego, CA, USA). Anti‐phospho‐CREB (Ser133) and anti‐CREB were purchased from Cell Signaling Technology (Danvers, MA, USA), anti‐Epac2 from Proteintech (Chicago, IL, USA), anti‐Rap1 from Millipore, peroxidase‐conjugated anti‐rabbit IgG and anti‐mouse IgG from Jackson Immuno Research (West Grove, PA, USA), anti‐glucokinase was produced in‐house
19 (link), and Alexa Fluor 488‐conjugated anti‐rabbit IgG (H + L) was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
19 (link), and Alexa Fluor 488‐conjugated anti‐rabbit IgG (H + L) was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
4
Doxorubicin Cardiotoxicity Evaluation
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Dox (2 mg/mL) and dexrazoxane were obtained from ACCORD (central pharmacy of Institut Gustave Roussy, Villejuif, France) and Sigma (St Quentin Fallavier, France), respectively. The in vitro Dox exposure of isolated cardiomyocytes was of 1–10 µM in concordance with literature recommendation (Tokarska-Schlattner et al., 2006 (link)) and extended to match tumor cell lines exposure requirements.
8-(4-Chloro-phenylthio)-2’-O-methyladenosine-3’5’cyclic monophosphate (8-CPT), ESI-09, and ESI-05 were from Biolog Life Science Institute (Bremen, Germany). ZVAD-fmk was from Bachem (Bubendorf, Switzerland). EPAC1 specific inhibitor (R)-CE3F4 was provided by Dr Ambroise (CEA, Gif-sur-Yvette, France) (Courilleau et al., 2013 (link)).
8-(4-Chloro-phenylthio)-2’-O-methyladenosine-3’5’cyclic monophosphate (8-CPT), ESI-09, and ESI-05 were from Biolog Life Science Institute (Bremen, Germany). ZVAD-fmk was from Bachem (Bubendorf, Switzerland). EPAC1 specific inhibitor (R)-CE3F4 was provided by Dr Ambroise (CEA, Gif-sur-Yvette, France) (Courilleau et al., 2013 (link)).
5
Preparation of MK-4, H89, and ESI-05 Stock Solutions
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MK-4 was obtained from Nisshin Pharma Inc. (Tokyo, Japan) and dissolved in ethanol to obtain a stock solution (50 mM); it was then stored in the dark at −20 °C. H89 (Sigma-Aldrich, St. Louis, MO, USA) and ESI-05 (4-methylphenyl-2,4,6-trimethlyphenylsulfone; BIOLOG Life Science Institute, Bremen, Germany), which are inhibitors of PKA and Epac2, respectively, were dissolved in dimethyl sulfoxide (Sigma-Aldrich) to obtain stock solutions (10 mM) at −20 °C.
6
Calcium Signaling in Rat Ventricular Myocytes
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Wistar rats (150–200 g) were sacrificed in accordance with the UK Home Office Guidance on the Operation of Animals (Scientific Procedures) Act of 1986. ARVMs were isolated by collagenase digestion as described previously (61 (link)). ARVMs were perfused with solutions containing (mM) 113 NaCl; 5.4 KCl; 1 MgCl2; 1.0 CaCl2; 0.37 Na2HPO4; 5.5 glucose; and 5 HEPES, 20–22°C, pH 7.1. Changes in intracellular calcium ([Ca2+]i) were detected by loading myocytes with fluo-4 AM (5 μM) for 15 min at room temperature (20–22°C). A further 30 min was allowed for washout of fluo-4 AM and de-esterification.
Unless otherwise stated, all chemicals were obtained from Sigma Aldrich. The selective Epac2 inhibitors, ESI-05 (35 (link), 50 (link)) and HJC0350 (14 (link)), were obtained from Biolog and Bio-techne/Tocris, respectively. The GGT-1 inhibitor, GGTI-298 (22 (link)), is the ester prodrug form of the parent compound GGTI-297 and both were obtained from Bio-techne/Tocris. In some experiments, GGTI-297 was introduced via a patch pipette during whole-cell patch-clamp recording, while GGTI-298 was added via the bathing solution. The GGT-1 inhibitor, GGTI-2147, was obtained from Calbiochem. Active Rap1 pull-down and detection kits were obtained from ThermoFisher Scientific.
Unless otherwise stated, all chemicals were obtained from Sigma Aldrich. The selective Epac2 inhibitors, ESI-05 (35 (link), 50 (link)) and HJC0350 (14 (link)), were obtained from Biolog and Bio-techne/Tocris, respectively. The GGT-1 inhibitor, GGTI-298 (22 (link)), is the ester prodrug form of the parent compound GGTI-297 and both were obtained from Bio-techne/Tocris. In some experiments, GGTI-297 was introduced via a patch pipette during whole-cell patch-clamp recording, while GGTI-298 was added via the bathing solution. The GGT-1 inhibitor, GGTI-2147, was obtained from Calbiochem. Active Rap1 pull-down and detection kits were obtained from ThermoFisher Scientific.
7
Modulation of Ca2+ Signaling Pathways
8
Evaluation of Cellular Signaling Modulators
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Cell culture reagents were obtained from Invitrogen (Darmstadt, Germany). The anti-pERK-1/2 (E-4) and the anti-Rap-1a (sc-1482) antiserum was from Santa Cruz (Heidelberg, Germany). The pCREB-Ser-133 antibody was purchased from cell signaling (Leiden, the Netherlands). The anti-tubulin (clone 6-11B-1) and the peroxidase-conjugated anti-mouse or anti-rabbit antibody, both raised in goat, from Bio-Rad (München, Germany). The firefly luciferase substrate was from Promega (Mannheim, Germany). rp-Br-cAMPs and γ-MSH were from SigmaAldrich (Deisenhofen, Germany). ESI-09, ESI-05 and HJC-0197 were from Biolog (Bremen, Germany), A-812511 from abcam (Cambridge, UK) and PD-184,352, HS-024 or NPY from Tocris (Bristol, UK). α-MSH was from Biotrend (Cologne, Germany) and KT-5720 from Enzo life science (Lörrach, Germany).
9
Excitatory Synaptic Transmission Assay
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For mEPSC recordings, TTX (Sigma) was added to the bath solution to only measure excitatory spontaneous release. For investigation of the cAMP-GluA3-dependent pathway, we used FSK (Sigma), H89 (Tocris), KT5720 (Sigma), ESI-05 (BioLog), and 8-CPT-2Me-cAMP (Tocris Bioscience). To obtain a monophasic time decay of the AMPA-evoked responses in outside-out patches, we added PEPA (Tocris bioscience) and cyclothiazide (Tocris bioscience).
10
Doxorubicin Cardiotoxicity Evaluation
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Dox (2 mg/mL) and dexrazoxane were obtained from ACCORD (central pharmacy of Institut Gustave Roussy, Villejuif, France) and Sigma (St Quentin Fallavier, France), respectively. The in vitro Dox exposure of isolated cardiomyocytes was of 1–10 µM in concordance with literature recommendation (Tokarska-Schlattner et al., 2006 (link)) and extended to match tumor cell lines exposure requirements.
8-(4-Chloro-phenylthio)-2’-O-methyladenosine-3’5’cyclic monophosphate (8-CPT), ESI-09, and ESI-05 were from Biolog Life Science Institute (Bremen, Germany). ZVAD-fmk was from Bachem (Bubendorf, Switzerland). EPAC1 specific inhibitor (R)-CE3F4 was provided by Dr Ambroise (CEA, Gif-sur-Yvette, France) (Courilleau et al., 2013 (link)).
8-(4-Chloro-phenylthio)-2’-O-methyladenosine-3’5’cyclic monophosphate (8-CPT), ESI-09, and ESI-05 were from Biolog Life Science Institute (Bremen, Germany). ZVAD-fmk was from Bachem (Bubendorf, Switzerland). EPAC1 specific inhibitor (R)-CE3F4 was provided by Dr Ambroise (CEA, Gif-sur-Yvette, France) (Courilleau et al., 2013 (link)).
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