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Amiloride
Amiloride
Amiloride is a potassium-sparing diuretic medication used to treat hypertension, heart failure, and edema.
It works by inhibiting the reabsorption of sodium and water in the kidneys, leading to increased urine output.
Amiloride has also been investigated for its potential neuroprotective effects in neurological disorders.
Reserach on Amiloride is critical for understanding its mechanisms of action and optimizing its clinical applications.
Pubcomapre.ai is an innovative AI-driven platform that can enhance reproducibility and accuracy in Amiloride research by streamlining the process of locating protocols and leverging AI-driven comparisons to identify the best protocols and products.
This tool can help improve Amiloride studies and workflows.
It works by inhibiting the reabsorption of sodium and water in the kidneys, leading to increased urine output.
Amiloride has also been investigated for its potential neuroprotective effects in neurological disorders.
Reserach on Amiloride is critical for understanding its mechanisms of action and optimizing its clinical applications.
Pubcomapre.ai is an innovative AI-driven platform that can enhance reproducibility and accuracy in Amiloride research by streamlining the process of locating protocols and leverging AI-driven comparisons to identify the best protocols and products.
This tool can help improve Amiloride studies and workflows.
Most cited protocols related to «Amiloride»
Acids
alanylglutamate
Amiloride
Binding Sites
Crystallography
Electrons
glutamylalanine
glycylvaline
Helix (Snails)
Labyrinth Fenestration
Microtubule-Associated Proteins
phenylalanylglutamate
Protein Subunits
Vestibular Labyrinth
2-(dimethylaminostyryl)-1-ethylpyridinium
Amikacin
Amiloride
Aminoglycosides
ARID1A protein, human
Auditory Hair Cell
Cell Survival
Embryo
Fishes
Gentamicin
Hair
Investigational New Drugs
Kanamycin
Larva
Neomycin
Pharmaceutical Preparations
Streptomycin
Tobramycin
tricaine
Zebrafish
To measure macropinocytosis, serum-starved A431 cells grown on coverslips and placed in Chamlid chambers were incubated with 0.5 mg/ml TMR-dextran and, where noted, stimulated with 100–200 ng/ml EGF in the indicated buffer for 10 min at 37°C. Cells were washed and both DIC and red fluorescence images of live cells were acquired. Where indicated, the following inhibitors were used: 10 µM latrunculin B, 100 µM LY294002, 1 mM amiloride, or 10 µM HOE-694. In the case of latrunculin B and LY294002 the cells were preincubated with the inhibitors at 37°C for 30 min before EGF addition. Macropinocytosis was quantified as the number of cells containing macropinosomes in the cells outlining each island.
Endocytosis was assessed by incubating the cells with 50 µg/ml Alexa 546–conjugated transferrin in the indicated buffer for 15 min at 37°C, after which the cells were placed on ice and acid washed with 0.2 M acetic acid in 150 mM NaCl and PBS to remove exofacial fluorescence. The cells were then fixed and mounted on slides, and red fluorescence was imaged and quantified.
Endocytosis was assessed by incubating the cells with 50 µg/ml Alexa 546–conjugated transferrin in the indicated buffer for 15 min at 37°C, after which the cells were placed on ice and acid washed with 0.2 M acetic acid in 150 mM NaCl and PBS to remove exofacial fluorescence. The cells were then fixed and mounted on slides, and red fluorescence was imaged and quantified.
Acetic Acid
Acids
Amiloride
Buffers
Cells
Dextran
Endocytosis
Erythrocytes
Fluorescence
HOE 694
inhibitors
latrunculin B
LY 294002
Serum
Sodium Chloride
Transferrin
SKOV3 exosomes were labeled with carboxyfluoresceine diacetate succinimidyl-ester (CFSE) (Invitrogen) as previously described [12 (link)]. Briefly, exosomes (20 μg) collected after a 100,000 × g ultracentrifugation were incubated with 7.5 μM CFSE for 30 min at 37°C in a final volume of 200 μl PBS containing 0.5% BSA. Labeled exosomes (Exos-CFSE) were 65-fold diluted with DMEM supplemented with 10% vesicles-free fetal calf serum and pelleted by ultracentrifugation for 16 h at 10,0000 × g, 12°C. Exos-CFSE were resuspended in DMEM and incubated with SKOV3 cells at 37 or 4°C.
When indicated Exos-CFSE or cells were treated for 30 min with 100 μg/ml proteinase K, or for 2 h with 15 mU neuraminidase from V. cholerae or from A. urefaciens (Roche), before uptake. SKOV3 cells were also incubated, 30 min prior to and during uptake, with the inhibitors 10 μg/ml chlorpromazine, 5 μg/ml cytochalasin D, 50 μM 5-ethyl-N-isopropyl amiloride (EIPA) or 2% methyl-beta-cyclodextrin, or with 150 mM of the monosaccharides D-glucose, D-galactose, α-L-fucose, α-D-mannose, D-N-acetylglucosamine, and the disaccharide β-lactose (Sigma).
Uptake assays were always performed in the presence of the compounds and analyzed after 2 or 4 h by immunofluorescence microscopy or flow cytometry.
When indicated Exos-CFSE or cells were treated for 30 min with 100 μg/ml proteinase K, or for 2 h with 15 mU neuraminidase from V. cholerae or from A. urefaciens (Roche), before uptake. SKOV3 cells were also incubated, 30 min prior to and during uptake, with the inhibitors 10 μg/ml chlorpromazine, 5 μg/ml cytochalasin D, 50 μM 5-ethyl-N-isopropyl amiloride (EIPA) or 2% methyl-beta-cyclodextrin, or with 150 mM of the monosaccharides D-glucose, D-galactose, α-L-fucose, α-D-mannose, D-N-acetylglucosamine, and the disaccharide β-lactose (Sigma).
Uptake assays were always performed in the presence of the compounds and analyzed after 2 or 4 h by immunofluorescence microscopy or flow cytometry.
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Acetylglucosamine
Amiloride
Biological Assay
Cells
Chlorpromazine
Cytochalasin D
Disaccharides
Endopeptidase K
Esters
Exosomes
Fetal Bovine Serum
Flow Cytometry
Fucose
Galactose
Glucose
Immunofluorescence Microscopy
inhibitors
Lactose
Mannose
methyl-beta-cyclodextrin
Monosaccharides
Neuraminidase
Ultracentrifugation
Vibrio cholerae
Amiloride
Aminoglycosides
Auditory Hair Cell
Embryo
Fishes
Gentamicin
Ions
Larva
Neomycin
Pharmaceutical Preparations
Tissues
Zebrafish
Most recents protocols related to «Amiloride»
For in vitro studies, OHP (Sigma–Aldrich Inc., Milano, Italy), cariporide (Sigma–Aldrich Inc., Italy), amiloride (Sigma–Aldrich Inc., Italy), 5-FU (Sigma–Aldrich Inc., Italy), FK506 (Bio-Techne, Milano, Italy), nigericin (Life Technologies, Monza, Italy) and valinomycin (Life Technologies, Monza, Italy) were used. These compounds, with the exception of OHP (reconstituted in 100% water) and FK506 (reconstituted in 100% ethanol), were dissolved in 100% dimethyl sulfoxide (DMSO) and stored at − 20 °C, according to the manufacturers’ specifications. For in vivo studies, OHP (Accord, Healthcare Italia) and 5-FU (Accord, Healthcare Italia) compounds were used from stock solutions to achieve the final concentrations. For each experiment, working concentrations of these drugs were freshly prepared by diluting them in their relative vehicle.
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Amiloride
cariporide
Ethanol
FK-506
Nigericin
Pharmaceutical Preparations
Sulfoxide, Dimethyl
Valinomycin
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5-(6)-carboxyfluorescein diacetate succinimidyl ester
Amiloride
Biological Assay
carboxyfluorescein
Cells
Cell Survival
Chlorpromazine
Cytotoxin
Esters
Filipin
Flow Cytometry
Fluorescent Dyes
Homo sapiens
inhibitors
Mammals
Obesity
Omeprazole
Psychological Inhibition
The cells were seeded on the glass bottom of dedicated confocal dishes (NEST) and cultured for 24 h. Hoechst 33342 (Sigma) and CellMask Green Plasma Membrane Stain (Invitrogen), whose fluorescent excitation/emission maxima were 346/460 nm and 522/535 nm separately, were used to label cell nucleus and the plasma membrane of ECs. The working solutions with indicated final concentrations of 5‐(N‐ethyl‐N‐isopropyl) amiloride (Selleckchem), CPZ HCl (Selleckchem), methyl‐β‐cyclodextrin (MβCD, Selleckchem), cytochalasin D (Cyto D, Selleckchem), and NOC (Selleckchem) were prepared by diluting the stock solutions with ECM. ECs were pretreated with inhibitor at 37 °C for 1 h before the attachment of sEVs and the corresponding inhibitor was maintained throughout the preparation stage. For static imaging, ECs were incubated with labeled sEVs (30 µg mL−1) at indicated temperature for indicated time. Then, the dish was washed gently by PBS for three times and fixed with 4% paraformaldehyde solution for 15 min at room temperature. Before observation, the dish was washed gently for three times with PBS. For live cell tracking, 50 nK LysoTracker Green (Meiunbio) was used to specifically label acidic endosomes according to research purpose. ECs were then incubated with labeled sEVs (30 µg mL−1) at 37 °C for 10 min. After washed gently with PBS, the dish was immobilized in an online live cell culture chamber (INUBG2‐PI, TOKAI HIT). Time‐lapse live‐cell images were acquired with a spinning‐disk confocal microscope system (the Andor Revolution XD) which was equipped with an invert microscope (IX 81, Olympus). A Nipkow disk type confocal unit (CSU 22, Yokogawa), an Emission filter wheel (Sutter Instruments), and an electron multiplying charge‐coupled device (EMCCD) (Andor iXon Ultra 897) were equipped to support the imaging speed. Diode pumped solid state (DPSS) lasers at 405, 488, 561, and 640 nm were used to excite Hochest 33342/Alexa Fluor 405, CellMask Green/FAM/LysoTracker Green, PKH26, and QDs 705, respectively.
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Acids
Amiloride
Cell Nucleus
Cells
Cells, Immobilized
Cyclodextrins
Cytochalasin D
Electrons
Endosomes
HOE 33342
Hyperostosis, Diffuse Idiopathic Skeletal
Lasers, Semiconductor
LysoTracker
Medical Devices
Microscopy
Microscopy, Confocal
paraform
PKH 26
Plasma Membrane
Stains
MDA‐MB‐231 cells were inoculated in the lower chamber of Transwell (pore size, 0.4 µm) at a density of 5 × 104 per well. Then amNR (0.1 mL, [DOX] = 50 µg mL−1) was added to the upper chamber. Meanwhile, the medium of the lower chamber was replaced with 0.5 mL of DMEM medium (pH 7.4 or 6.5). A cylindrical permanent magnet was placed below the Transwell plate for the magnetically actuated treatment. After 4 h of incubation, these samples were harvested for flow cytometry analysis. In addition, cell internalization of amNR was further visualized by CLSM. Moreover, Nuclear fast red and Prussian blue double staining of MDA‐MB‐231 cells was also performed to evaluate tumor cell uptake. To study the uptake mechanism, MDA‐MB‐231 cells were pre‐incubated with various inhibitors including Methyl‐β‐cyclodextrin (5.0 µm ), chlorpromazine (10 µg mL−1), and amiloride (50 µm ). The incubation was at 37 °C and for 1 h prior to treatment with amNR or amNR + pH 6.5 for another 4 h. Subsequently, the cells were washed twice and trypsinized for flow cytometry analyses.
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Amiloride
Cells
Chlorpromazine
Cyclodextrins
Flow Cytometry
inhibitors
MDA-MB-231 Cells
Neoplasms
We loaded OVA-FITC into silica nanoparticles and measured the average intracellular fluorescence intensity by FACS analysis to quantitatively detect the uptake of different silica nanoparticles. RAW264.7 cells (3.0 × 105 cells) were precultured in 12-well plates for 12 h and then incubated with free OVA-FITC plus IMQ, OVA-FITC-IMQ-VMSNs/MSNs (25 μg/mL OVA-FITC, 5 μg/mL IMQ) for 15 min, 30 min, 1 h, 2 h, 4 h and 8 h. After that, the cells were washed with PBS for more than 3 times to terminate ingestion and measured average intracellular fluorescence intensity and OVA-FITC positive RAW264.7 cells by FACS analysis. Absorption of VMSNs by cells 4 h after incubation were observed using CLSM under the same conditions.
With the aim of investigating the behavior of internalized silica nanoparticles, we used CLSM to observe the location of silica nanoparticles. RAW264.7 cells were incubated with free OVA-FITC plus IMQ, OVA-FITC-IMQ-VMSNs/MSNs (25 μg/mL OVA-FITC, 5 μg/mL IMQ) for 12 h after well plate growth. To determine the degree of co-localization of OVA and lysosomes, the images were treated with a CLSM after the cell nucleus was labeled by Hoechst and lysosomes stained by Lyso-Tracker Red.
To further investigate the cell uptake mechanism, RAW264.7 cells (3.0 × 105 cells) were incubated in 24-well plates for 12 h. Before adding silica nanoparticles, the cells were incubated with the corresponding mechanism inhibitors for 1h, that is, chlorpromazine (CPZ, 20 μg/mL) was used to inhibit clathrin-mediated endocytosis, genistein (Geni, 2 μg/mL) was employed to suppress the caveolae-mediated endocytosis, and amiloride (Amil, 15 μg/mL) was served as the macropinocytosis inhibitor. The cells were then cultured with the OVA-IMQ-VMSNs/MSNs corresponding inhibitors for 6 h. Results were expressed as percentage uptake in the control group incubated with OVQ-IMQ-VMSNs/MSNs without inhibitors.
With the aim of investigating the behavior of internalized silica nanoparticles, we used CLSM to observe the location of silica nanoparticles. RAW264.7 cells were incubated with free OVA-FITC plus IMQ, OVA-FITC-IMQ-VMSNs/MSNs (25 μg/mL OVA-FITC, 5 μg/mL IMQ) for 12 h after well plate growth. To determine the degree of co-localization of OVA and lysosomes, the images were treated with a CLSM after the cell nucleus was labeled by Hoechst and lysosomes stained by Lyso-Tracker Red.
To further investigate the cell uptake mechanism, RAW264.7 cells (3.0 × 105 cells) were incubated in 24-well plates for 12 h. Before adding silica nanoparticles, the cells were incubated with the corresponding mechanism inhibitors for 1h, that is, chlorpromazine (CPZ, 20 μg/mL) was used to inhibit clathrin-mediated endocytosis, genistein (Geni, 2 μg/mL) was employed to suppress the caveolae-mediated endocytosis, and amiloride (Amil, 15 μg/mL) was served as the macropinocytosis inhibitor. The cells were then cultured with the OVA-IMQ-VMSNs/MSNs corresponding inhibitors for 6 h. Results were expressed as percentage uptake in the control group incubated with OVQ-IMQ-VMSNs/MSNs without inhibitors.
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Amiloride
Cardiac Arrest
Caveolae
Cell Nucleus
Cells
Clathrin
Endocytosis
Fluorescein-5-isothiocyanate
Fluorescence
Genistein
inhibitors
Lysosomes
LysoTracker
MSN protein, human
Protoplasm
RAW 264.7 Cells
Silicon Dioxide
Top products related to «Amiloride»
Sourced in United States, Germany, Italy, Sao Tome and Principe, United Kingdom, Macao, China, Denmark
Amiloride is a laboratory product manufactured by Merck Group. It is a small molecule compound primarily used as a research tool in scientific investigations. Amiloride functions as a potassium-sparing diuretic, inhibiting the sodium-hydrogen exchanger and the epithelial sodium channel.
Sourced in United States, United Kingdom, China, Germany
Chlorpromazine is a pharmaceutical compound used as a laboratory reagent. It is a white crystalline solid that is soluble in water and organic solvents. Chlorpromazine is commonly used in research and laboratory settings as a reference standard or for various analytical purposes.
Sourced in United States, Germany
5-(N-ethyl-N-isopropyl) amiloride (EIPA) is a chemical compound that functions as a selective inhibitor of the Na+/H+ exchanger (NHE). It is commonly used as a research tool in laboratory settings to study the role of NHE in various biological processes.
Sourced in United States
Ussing chambers are specialized laboratory equipment used to measure the transepithelial transport of ions, solutes, and water across epithelial cell layers. They provide a controlled environment for the study of physiological processes in biological membranes.
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Forskolin is a lab equipment product manufactured by Merck Group. It is a compound derived from the roots of the Coleus forskohlii plant. Forskolin is used as a tool for research purposes in the laboratory setting.
Sourced in United States, Germany, Sao Tome and Principe, United Kingdom, Israel
Dynasore is a small molecule compound used in laboratory research. It functions as a dynamin inhibitor, a protein involved in processes such as endocytosis. The core function of Dynasore is to inhibit the activity of dynamin, which is important for various cellular processes.
Sourced in United States, Germany, China, France, United Kingdom, Sao Tome and Principe, Italy, Australia, Poland
Genistein is a lab equipment product from Merck Group. It is a naturally occurring isoflavone compound found in various plants. Genistein can be used as a research tool in various scientific applications.
Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal
Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Nystatin is an antifungal medication used in the laboratory setting. It is a polyene macrolide antibiotic that functions by disrupting the cell membrane of fungal cells, leading to their death. Nystatin is commonly used for the treatment and prevention of fungal infections in in vitro experiments and laboratory procedures.
Sourced in United States, Germany, France
CFTRinh-172 is a laboratory reagent that functions as a selective inhibitor of the cystic fibrosis transmembrane conductance regulator (CFTR) protein. It is used in research applications to study the role of CFTR in various cellular processes.
More about "Amiloride"
Amiloride is a potassium-sparing diuretic medication that is widely used to treat conditions like hypertension, heart failure, and edema.
This drug works by inhibiting the reabsorption of sodium and water in the kidneys, leading to increased urine output.
Amiloride has also shown promise for its potential neuroprotective effects in various neurological disorders.
Research on Amiloride is crucial for understanding its mechanisms of action and optimizing its clinical applications.
Pubcomapre.ai, an innovative AI-driven platform, can enhance reproducibility and accuracy in Amiloride research by streamlining the process of locating protocols and leverging AI-driven comparisons to identify the best protocols and products.
This tool can help improve Amiloride studies and workflows.
Other related compounds like Chlorpromazine, 5-(N-ethyl-N-isopropyl) amiloride (EIPA), and Forskolin have also been studied in the context of Amiloride research.
Techniques such as Ussing chambers, Dynasore, Genistein, and the use of FBS and Nystatin have been employed to investigate the effects of these compounds.
The CFTR inhibitor CFTRinh-172 has also been utilized in Amiloride research to understand its mechanisms of action.
This drug works by inhibiting the reabsorption of sodium and water in the kidneys, leading to increased urine output.
Amiloride has also shown promise for its potential neuroprotective effects in various neurological disorders.
Research on Amiloride is crucial for understanding its mechanisms of action and optimizing its clinical applications.
Pubcomapre.ai, an innovative AI-driven platform, can enhance reproducibility and accuracy in Amiloride research by streamlining the process of locating protocols and leverging AI-driven comparisons to identify the best protocols and products.
This tool can help improve Amiloride studies and workflows.
Other related compounds like Chlorpromazine, 5-(N-ethyl-N-isopropyl) amiloride (EIPA), and Forskolin have also been studied in the context of Amiloride research.
Techniques such as Ussing chambers, Dynasore, Genistein, and the use of FBS and Nystatin have been employed to investigate the effects of these compounds.
The CFTR inhibitor CFTRinh-172 has also been utilized in Amiloride research to understand its mechanisms of action.