6-Bnz-cAMP-AM, Me-cAMP-AM (8-pCPT-2′-O-Me-cAMP-AM), and ESI-09 were purchased from Biolog (Germany). Ryanodine, SKF-96365, and Xestospongin C were purchased from MilliporeSigma (Burlington, MA). Bt3-Ins(1,3,5)P3/AM was purchased from SiChem. ATP, 4-CMC (4-Chloro-m-cresol), H-89, and thapsigargin were from Sigma-Aldrich (St. Louis, MO).
Esi 09
Manufactured by Biolog
Sourced in Germany
The ESI-09 is a laboratory equipment used for electrospray ionization (ESI) in mass spectrometry applications. It provides a reliable and consistent method for converting liquid samples into a fine mist of charged droplets, which can then be analyzed by a mass spectrometer. The core function of the ESI-09 is to facilitate the efficient transfer of analytes from the liquid phase to the gas phase, enabling their subsequent detection and identification.
Automatically generated - may contain errors
Lab products found in correlation
Esi 05, by Biolog (2 mentions)
Hjc0197, by Biolog (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
6 bnz camp am, by Biolog (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Thapsigargin, by Merck Group (1 mentions)
L nna, by Merck Group (1 mentions)
Mdl 12 330a, by Merck Group (1 mentions)
Pierce ecl solution, by Thermo Fisher Scientific (1 mentions)
Ec basal medium plus supplement pack, by PromoCell (1 mentions)
Rac1 activation assay kit, by Cytoskeleton (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Histamine dihydrochloride, by Merck Group (1 mentions)
Forskolin, by Merck Group (1 mentions)
8 pcpt 2 o me camp, by Biolog (1 mentions)
6 bnz camp, by Biolog (1 mentions)
Rp camps, by Biolog (1 mentions)
Sq22536, by Merck Group (1 mentions)
Butaprost free acid, by Cayman Chemical (1 mentions)
Rp 8 cpt camps, by Biolog (1 mentions)
9 protocols using esi 09
1
Modulation of Intracellular Signaling Pathways
2
Endothelial Cell Barrier Permeability Assay
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HRP‐conjugated anti‐mouse IgG, and rabbit IgG antibodies were purchased from Amersham Biosciences (Heidelberg, Germany); human thrombin from Behring (Marburg, Germany); anti VE‐cadherin (clone TEA 1, mouse IgG) from Beckman Coulter (Krefeld, Germany); ESI‐09 from Biolog (Bremen, Germany); benzonase, forskolin, myristoylated PKI, and Y‐27632, from Calbiochem (Darmstadt, Germany); anti‐phospho‐MLC and anti‐GAPDH antibodies from Cell signaling (Danvers, MA); Rac1 activation assay kit from cytoskeleton (Denver); Pierce® ECL solution from Fischer Scientific (Niederlassung Nidderau, Germany); Alexa‐Flour labelled anti‐mouse IgG and anti‐rabbit IgG antibodies from Invitrogen (Karlsruhe, Germany); anti‐Rac1‐GTP antibody from NewEast Biosciences (King of Prussia, PA); EC basal medium plus supplement pack from PromoCell (Heidelberg, Germany); Complete® protease inhibitor cocktail from Roche (Mannheim, Germany); 3‐isobutyl‐1‐methylxanthine (IBMX), Phalloidin‐TRITC, L‐NAME, L‐NNA, MDL‐12 330A, and anti‐vinculin (clone hVIN‐1, mouse IgG) from Sigma (Steinheim, Germany); anti‐phospho‐MYPT1 (Thr850) from Merck Millipore (Schwalbach, Germany), and Transwell polycarbonate membrane filters (24‐mm round) were from Greiner Bio‐One (Frickenhausen, Germany). All other chemicals were of the best available quality, usually analytical grade.
3
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).
4
Reagents Used in Calcium Signaling Experiments
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H89, NKH 477, 8-Br-cAMP, and 8-Br-cGMP were from R&D Systems (Abingdon, Oxford, UK). Sp-cAMPS, 6-Bnz-cAMP, 8-pCPT-2′-O-Me-cAMP, Rp-cAMPS, Rp-8-CPT-cAMPS, ESI-09, and HJC0197 were from Biolog (Bremen, Germany). Ionomycin, SQ 22536, DDA and myristoylated-PKA inhibitor peptide (PKI) were from Merck-Millipore (Watford, UK). A membrane-permeant peptide inhibitor of A kinase-anchoring proteins (AKAPs) [stearated Ht31 AKAP inhibitor peptide (st-Ht31)] and its proline-modified inactive form (st-Ht31P) were from Promega (Southampton, UK). Thapsigargin was from Alomone Laboratories (Jerusalem, Israel). PAR1 peptide, histamine dihydrochloride, forskolin, IBMX, and PGE2 were from Sigma-Aldrich (Welwyn Garden City, UK). Butaprost (free acid) and L902,688 were from Cayman Chemicals (Ann Arbor, MI). Membrane-permeant caged IP3 (ci-IP3PM) was from SiChem (Bremen, Germany). [2,8-3H] adenine ci-IP3PM, d -2,3-O-isopropylidene-6-O-(2-nitro-4,5-dimethoxy)benzyl-myo-inositol 1,4,5-trisphosphate-hexakis(propionoxymethyl) ester was from Perkin Elmer (Seer Green, Bucks, UK). Fluo-8 was from Stratech Scientific Ltd (Newmarket, Suffolk, UK). Other reagents were from Sigma-Aldrich, sources specified previously (Pantazaka et al., 2013 (link)) or identified in this section.
5
Modulation of cAMP Signaling Pathways
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Sp-adenosine 3′,5′-cyclic monophosphorothioate triethylammonium salt hydrate (Sp-cAMPS) and Rp-adenosine 3′,5′-cyclic monophosphorothioate triethylammonium salt hydrate (Rp-cAMPS) were applied at 100 μM. ESI-09 (membrane-permeant inhibitor of Epac 1 and Epac 2, BIOLog) was applied at 10, 30 or 100 μM. Protein Kinase A inhibitor fragment 14–22 (PKI) was applied at 20 μM. N6-phenyladenosine- 3′,5′- cyclic monophosphate (6-phe-cAMP) was applied at 100 μM. 8-(4-chlorophenylthio)-2′-O-methyladenosine 3′,5′-cyclic monophosphate monosodium (8-cpt-2Me-cAMP) was applied at 200 μM.
6
Characterization of cAMP Analogues and Inhibitors
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The cell‐permeable cAMP analogues: 8‐Br (8‐bromoadenosine‐3′,5′‐cyclic monophosphate, acetoxymethyl ester); 6‐Bnz (N6‐benzoyladenosine‐3′,5′‐cyclic monophosphate, acetoxymethyl ester; CPT (8‐(4‐chlorophenylthio)‐2′‐O‐methyladenosine‐3′,5′‐cyclic monophosphate, acetoxymethyl ester); and the Epac inhibitor, ESI‐09 (3‐[5‐(tert.‐butyl)isoxazol‐3‐yl]‐2‐[2‐(3‐chlorophenyl)hydrazono]‐3‐oxopropanenitrile) (Almahariq et al., 2013 ) were purchased from BioLog Life Science Institute (Bremen, Germany). The PKA inhibitor 14–22 amide (PKI), cell‐permeable, myristoylated was purchased from Merck Millipore (Watford, UK). Other biochemicals were purchased from Sigma (Gillingham, UK), including the PKA inhibitor H‐89 and the PKC inhibitor chelerythrine. General chemicals were purchased from Fischer Scientific (Loughborough, UK) or VWR‐Jencons (Lutterworth, UK).
7
Biochemical Reagents Characterization
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
Investigating LPS-Induced Inflammatory Responses
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Escherichia coli LPS (O55:B5), 8-bromoadenosine 3′,5′-cyclic
monophosphate (8-bromo-cAMP),
3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Giemsa
stain, rolipram, and roflumilast were purchased from Sigma-Aldrich (St Louis,
MO, USA), murine monocyte chemoattractant protein-1 (MCP-1) is from PeproTech
(Rocky Hill, NJ, USA), and the Epac inhibitor ESI-09 and PKA inhibitor
Rp-8-CPT-cAMPS were from BioLog Life Science Institute (Bremen, Germany). The
DMEM medium base, FBS, and horse serum were obtained from Thermo Fisher
Scientific (Waltham, MA, USA).
monophosphate (8-bromo-cAMP),
3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Giemsa
stain, rolipram, and roflumilast were purchased from Sigma-Aldrich (St Louis,
MO, USA), murine monocyte chemoattractant protein-1 (MCP-1) is from PeproTech
(Rocky Hill, NJ, USA), and the Epac inhibitor ESI-09 and PKA inhibitor
Rp-8-CPT-cAMPS were from BioLog Life Science Institute (Bremen, Germany). The
DMEM medium base, FBS, and horse serum were obtained from Thermo Fisher
Scientific (Waltham, MA, USA).
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