N-(p-amylcinnamoyl)anthranilic acid (ACA), maintained as a 50 mM stock solution in dimethylsulfoxide (DMSO), 2-aminoethoxydiphenyl borate (2-APB; 75 mM stock solution in DMSO), aristolochic acid (75 mM stock solution in DMSO) and 3-methyladenine (3-MA; 50 mM stock solution in DMSO) were purchased from Sigma (St. Louis, MO, USA). N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG; 0.5 M stock solution in DMSO) was purchased from AccuStandard (New Haven, CT, USA). Doxorubicin hydrochloride and tamoxifen citrate were purchased from Thermo Scientific (Waltham, MA, USA). Q-VD-OPh was purchased from R&D Systems (Minneapolis, MN, USA) and maintained as a 50 mM stock solution in DMSO. Propidium iodide (PI) (10 mg/ml solution) was purchased from Thermo Scientific. ApoScreen Annexin V-fluorescein isothiocyanate (FITC) was purchased from Southern Biotech (Birmingham, AL, USA).
N p amylcinnamoyl anthranilic acid aca
Manufactured by Merck Group
Sourced in United States
N-(p-amylcinnamoyl)anthranilic acid (ACA) is a chemical compound that can be used as a laboratory reagent. It is a carboxylic acid derivative with a specific molecular structure. The core function of ACA is to serve as a chemical tool for research and analysis purposes, without further interpretation of its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Aristolochic acid, by Merck Group (1 mentions)
Tamoxifen citrate, by Thermo Fisher Scientific (1 mentions)
2 aminoethoxydiphenyl borate 2 apb, by Merck Group (1 mentions)
Doxorubicin hydrochloride, by Thermo Fisher Scientific (1 mentions)
Q vd oph, by R&D Systems (1 mentions)
3 methyladenine 3 ma, by Merck Group (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Suramin, by Merck Group (1 mentions)
Apyrase, by Merck Group (1 mentions)
Dihydrorhodamine 123 dhr 123, by Thermo Fisher Scientific (1 mentions)
N acetyl leu glu his asp 7 amino 4 methylcoumarin ac lehd amc, by Bachem (1 mentions)
Icilin, by Bio-Techne (1 mentions)
L menthol, by Merck Group (1 mentions)
Amtb hydrate, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Cytotox 96 non radioactive cytotoxicity assay, by Promega (1 mentions)
Fetal bovine serum (fbs), by Lonza (1 mentions)
Ecl detection system, by Thermo Fisher Scientific (1 mentions)
Anticleaved caspase 3antibody, by Affinity Biosciences (1 mentions)
5 protocols using n p amylcinnamoyl anthranilic acid aca
1
Chemical Compounds for Cell Biology
2
Calcium Imaging Protocol for L-menthol
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L-menthol was purchased from Sigma-Aldrich and dissolved in ethanol (100%) at 1 M concentration. The compound was further diluted with buffer solution used for Ca2+ imaging experiments that contained (in mM): NaCl 138, Na2PO4 8, CaCl2 2, MgCl2 0.5, KCl 2.7, KH2PO4 1.6; pH 7.4. In the low Ca2+ solution, CaCl2 was replaced with 10 mM EGTA. The final concentration of the solvents were < 0.1% in all experimental solutions. At these concentrations the solvents did not affect/modify the evoked Ca2+ responses in control experiments (data not shown). N-(p-amylcinnamoyl)-anthranilic acid (ACA), N-methyl-D glutamine (NMDG), apyrase, AMTB-hydrate (AMTB), suramin, N-acetyl-Asp-Glu-Val-Asp-7-amino-4-methylcoumarin (ACDEVD-AMC), penicillin-streptomycin and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, MO, USA). Dihydrorhodamine-123 (DHR-123) was from Molecular Probes (Eugene, OR, USA). N-acetyl-Leu-Glu-His-Asp-7-amino-4-methylcoumarin (AC-LEHD-AMC) was purchased from Bachem (Bubendorf, Switzerland). Icilin, Capsazepine (CapZ) and BCTC were obtained from Tocris Bioscience (Bristol, UK). All reagents were of analytical grade.
3
WNV Infection Assay with Inhibitors
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Cells were infected with WNVKUN strain MRM61C at a multiplicity of infection (m.o.i.) of 0.5, 2 or 3 as has been described previously (34). Vero and BHK cells (both from ATCC) were maintained in DMEM supplemented with 5% FBS (Lonza, Basel, Switzerland) at 37°C with 5% CO2. Vero, HEK-293T and LN-18 cells for gene silencing experiments and enzyme activity measurements (cPLA2 assay kit) were maintained in EMEM, supplemented with 1mM non-essential amino acids and 10% FBS. C6/36 mosquito cells were maintained in L-15 (Leibovitz) media buffered by phosphates and supplemented with 10% FBS at 28C. Chemicals C75 (final concentration of 30μM), Palmityl trifluoromethyl ketone (PACOCF3; final concentration of 15μM), N-(p-Amylcinnamoyl)anthranilic acid (ACA; final concentration of 20μM) and N-Ethylmaleimide (NME; final concentration of 100μM) were all purchased from Sigma. The lyso-PChol mix (final concentration of 1.5μM of 500nM 18:0, 500nM 18:1 and 500nM 20:0 lyso-PChol) and fluorophore tagged lyso-PChol (lyso-PChol488; final concentration of 5μM or 20μM) were purchased from Avanti Polar Lipids Inc. Cytotoxicity was assessed by serial dilution of each chemical on Vero cells and viability assessed with the CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega).
4
TRPM2 Modulation in Cancer Cell Apoptosis
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We used the following reagents and antibodies: RPMI-1640 medium and fetal bovine serum
(Biological Industries); anti-TRPM2 antibody (#HPA035260, Sigma); anticleaved caspase-3
antibody (#AF7022, Affinity Biosciences); anti-β-tubulin antibody (#AF7011, Affinity
Biosciences); horseradish peroxidase (HRP)-conjugated antirabbit (#S0001, Affinity
Biosciences); electrochemiluminescence (ECL) detection system (#WP20005, Thermo Fisher
Scientific, Pittsburgh); Cell Counting Kit-8 (CCK-8) (#A311-01, Vazyme Biotech); ROS assay
kit (#S0033S, Beyotime Biotech); N-(p-amylcinnamoyl) anthranilic acid (ACA) (#A8486-5MG,
Sigma); H2O2 (#HX0636, Sigma); and 5-FU (#F6627-1G, Sigma). Specific
small interfering (si)RNAs for human TRPM2 (5′-GUCUCGGACAUCACUAUCUTT-3′, 5′-GGGAAGAUGUCUCAGCAGACG-3′)16 (link)
and scrambled siRNAs (5′-ACGCGUAACGCGGGAAUUU-3′, 5′-UUCUCCGAACGUGUCACGUTT-3′) were
designed and synthesized by Biomics Company.
(Biological Industries); anti-TRPM2 antibody (#HPA035260, Sigma); anticleaved caspase-3
antibody (#AF7022, Affinity Biosciences); anti-β-tubulin antibody (#AF7011, Affinity
Biosciences); horseradish peroxidase (HRP)-conjugated antirabbit (#S0001, Affinity
Biosciences); electrochemiluminescence (ECL) detection system (#WP20005, Thermo Fisher
Scientific, Pittsburgh); Cell Counting Kit-8 (CCK-8) (#A311-01, Vazyme Biotech); ROS assay
kit (#S0033S, Beyotime Biotech); N-(p-amylcinnamoyl) anthranilic acid (ACA) (#A8486-5MG,
Sigma); H2O2 (#HX0636, Sigma); and 5-FU (#F6627-1G, Sigma). Specific
small interfering (si)RNAs for human TRPM2 (5′-GUCUCGGACAUCACUAUCUTT-3′, 5′-GGGAAGAUGUCUCAGCAGACG-3′)16 (link)
and scrambled siRNAs (5′-ACGCGUAACGCGGGAAUUU-3′, 5′-UUCUCCGAACGUGUCACGUTT-3′) were
designed and synthesized by Biomics Company.
5
Whole-cell Patch-clamp Recordings of TRPM2 Currents
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Whole-cell recordings were carried out at room temperature using an Axopatch 200B amplifier. Patch electrodes with a resistance of 3–5 MΩ were fabricated from borosilicate glass (Sutter Instrument). The protocols for electrophysiological recordings were described in our previous studies (Luo et al., 2018 (link), 2019 (link)). Briefly, voltage ramps with 500 ms duration from-100 mV to +100 mV were applied every 5 s. The currents at-80 mV were denoted by circles in figures. The intracellular solution (ICS) of 500 μM ADPR contained: 147 mM NaCl, 1 mM MgCl2, 0.05 mM EGTA, 10 mM Hepes, and 500 μM ADPR (pH 7.4, adjusted with NaOH). The ICS of 50 mM Ca2+ contained: 50 mM CaCl2, 75 mM NaCl, 1 mM MgCl2, and 10 mM Hepes (pH 7.4, adjusted with NaOH). For all experiments, the standard extracellular solution (ECS) contained: 147 mM NaCl, 2 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM Hepes, and 13 mM glucose (pH 7.4, adjusted with NaOH). To confirm the TRPM2 channel currents, 20 mM N-(pamylcinnamoyl) anthranilic acid (ACA; Sigma) was applied at the end of each recording, and only the cells whose currents were completely inhibited by ACA were used for analysis.
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