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Probenecid

Probenecid is a uricosuric agent used to treat gout and hyperuricemia.
It works by inhibiting the reabsorption of uric acid in the kidneys, leading to increased urinary excretion of uric acid.
Probenecid may also be used to prevent or treat certain types of kidney stones.
It is commonly prescribed in combination with other medications, such as antibiotics, to enhance their effectiveness.
Probenecid is generally well-tolerated, but may cause side effects like gastrointestinal discomfort, rash, and headache in some patients.
Monitoring of uric acid levels and kidney function is recommended during treatment.
Probenecid is an important tool in the management of gout and related conditions, offering an effective way to lower uric acid levels and improve patient outcomes.

Most cited protocols related to «Probenecid»

1
Ligand-Protein Docking of Phytochemicals
Cited 29 times
The data set was prepared based on an extensive literature survey taking IC50 values of in-vitro enzyme inhibition assays against XO and HMGR by various secondary metabolites. Based on IC50 values, sixteen plant- and fungus-based secondary metabolites (Tables 1 and 2) were chosen for the ligand-protein docking study. The docking study was performed against commercial drugs such as atorvastatin, simvastatin, lovastatin, and pravastatin for HMGR. On the other hand, commercial drugs such as allopurinol, febuxostat, topiroxostat, and probenecid were used for molecular docking studies with XO. The structures of the ligand molecules and the control drugs of both enzymes were retrieved from the PubChem database [38 (link)] and verified from SciFinder. The structures were retrieved in SDF format and were converted to PDB and MOL2 format using Discovery Studio Visualizer 4.0 software. The structure and complete chemical properties, torsional energy, van der Waals potential energy, electrostatic energy, weight, log P, total polar surface area (TPSA), donor atoms, and acceptor atoms of the ligands were listed (Supplementary Table 4S) by the help of MOE Module [39 (link)].
Marahatha R., Basnet S., Bhattarai B.R., Budhathoki P., Aryal B., Adhikari B., Lamichhane G., Poudel D.K, & Parajuli N. (2021). Potential natural inhibitors of xanthine oxidase and HMG-CoA reductase in cholesterol regulation: in silico analysis. BMC Complementary Medicine and Therapies, 21, 1.
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Publication 2021
Allopurinol Atorvastatin chemical properties Electrostatics Enzyme Assays Enzymes Febuxostat Fungi Ligands Lovastatin Molecular Structure Pharmaceutical Preparations Plants Pravastatin Probenecid Proteins Psychological Inhibition Simvastatin Tissue Donors topiroxostat
2
Validating Self-Reported Gout Diagnosis in ARIC
Cited 9 times
ARIC is a population-based cohort study of 15,792 individuals recruited in 1987–89 from 4 US communities (Washington County, Maryland; Forsyth County, North Carolina; Jackson, Mississippi; Minneapolis, Minnesota; Figure 2). The institutional review board of the participating institutions (Johns Hopkins University, University of Mississippi, Wake Forest University, University of Minnesota, Baylor University, University of Texas, and University of North Carolina) approved the ARIC study protocol and study participants provided written informed consent. This study consisted of 1 baseline visit (visit 1) and 3 followup visits (visits 2, 3, and 4) administered 3 years apart. Importantly, there was a surveillance component to ARIC throughout the followup period. When a participant was hospitalized within the catchment area of the participating institutions, all of the corresponding discharge diagnoses were recorded.
Our study population consisted of men and women who self-reported their gout status at visit 4. Notably, at ARIC visit 4 each participant was asked, “Has a doctor ever told you that you had gout?”. Participants who answered “Yes” were considered to have a self-reported, physician-diagnosed case of gout. If a participant was recorded through surveillance as having a hospital discharge summary that listed an International Classification of Diseases (ICD)-9 code for gout (274.0, 274.1, 274.8, or 274.9), then they were considered to have gout based on this assignment of a gout discharge diagnosis. Therefore, if a participant attended visit 4 and the date of the gout-related hospitalization was prior to the visit 4 date, they were considered to be a gout case for the assessment of sensitivity. In addition, at each of the 4 ARIC visits, all medications used within the preceding month were recorded. We defined gout medications as colchicine, probenecid, and allopurinol. If a participant reported the use of any of these 3 medications at any study visit, they were considered to be a gout case. In our study, the gold standard for a diagnosis of gout was defined as either a hospital discharge diagnosis of gout or use of gout medication at any cohort visit. Although a prescription for these medications does not mean with absolute certainty that the ARIC participant has gout, in a random sample of 4 US communities, a prescription for colchicine, allopurinol, or probenecid is most likely issued to treat gout.
The gold standard gout definition was applied to all participants who attended visit 4. We calculated the sensitivity of a self-report of physician-diagnosed gout. Sensitivity was defined as the percentage of gold standard gout cases with a corresponding affirmative self-report of gout on the visit 4 questionnaire. Next, we conducted a stratified analysis for the sensitivity of a self-report of physician-diagnosed gout by sex, race, education, and hyperuricemia (serum urate level > 7.0 mg/dl at either visit 1 or 2) categories.
Additionally, we performed a sensitivity analysis to assess whether the sensitivity of a report of gout at visit 4 depended upon the definition of the gold standard. Specifically, we calculated sensitivity, separately, for participants with a hospital discharge diagnosis of gout as well as for those using gout medications. All analyses were performed in SAS, version 9.1 (SAS Institute, Cary, NC, USA).
McADAMS M.A., MAYNARD J.W., BAER A.N., KÖTTGEN A., CLIPP S., CORESH J, & GELBER A.C. (2010). Reliability and Sensitivity of the Self-report of Physician-diagnosed Gout in the Campaign Against Cancer and Heart Disease and the Atherosclerosis Risk in the Community Cohorts. The Journal of Rheumatology, 38(1), 135-141.
Publication 2010
Allopurinol Colchicine Diagnosis Ethics Committees, Research Forests Gold Gout Hospitalization Hypersensitivity Hyperuricemia Patient Discharge Pharmaceutical Preparations Physicians Probenecid Serum Urate Woman
3
Genetic determinants of gout in the UK Biobank
Cited 8 times
This research was conducted using the UK Biobank Resource (approval number 12611) [4 (link)]. Data from the first tranche of UK Biobank genotyping and imputation data were used for this analysis (made publicly available in May 2015). Inclusion criteria were European ethnicity, age 40–69 years and genome-wide genotypes available. Exclusion criteria were self-reported sex mismatch with genetic sex, genotyping quality control failure, related individuals, either a primary or secondary hospital diagnosis of kidney disease (International Classification of Diseases, Tenth Revision (ICD-10), codes I12, I13, N00-N05, N07, N11, N14, N17–N19, Q61, N25.0, Z49, Z94.0, Z99.2), participants aged 70 years and over, and those with kidney disease, because these are risk factors for secondary gout.
Gout definitions and combinations of these definitions were identified from previous epidemiological studies [1 (link), 3 (link), 5 (link)]. Self-report of gout was defined by reporting of gout by the participant at the time of the study interview. Hospital diagnosis of gout was defined by either primary or secondary hospital discharge coding for gout (ICD-10 code M10, including sub-codes). Use of ULT required self-report of being on any of allopurinol, febuxostat or sulphinpyrazone and not having a hospital diagnosis of leukaemia or lymphoma (ICD-10 codes C81–C96). Winnard-defined gout was hospital diagnosis of gout or gout-specific medication (ULT or colchicine) as reported by Winnard et al. [5 (link)]. For participants who did not meet any gout definitions, further exclusion criteria were corticosteroid use, non-steroidal anti-inflammatory drug use or probenecid use.
UK Biobank samples had been genotyped using an Axiom array (820,967 markers; Affymetrix, Santa Clara, CA, USA) and imputed to approximately 73.3 million single-nucleotide polymorphisms (SNPs) using SHAPEIT3 and IMPUTE2 with a combined UK10K and 1000 Genomes reference panel. Logistic regression of SNPs against gout as the outcome was performed, adjusting for age, sex, waist circumference, and ratio of waist circumference to height. We analysed 30 urate-associated SNPs reported by Köttgen et al. in the large (>140,000 European participants) Global Urate Genetics Consortium GWAS [1 (link)]. Data were reported on the basis of number of SNPs detected at both genome-wide significance (P < 5 × 10−8) and experiment-wide significance (P < 0.0017). CIs for proportions were calculated using the Wilson score method and www.openepi.com [6 ]. Heritability estimates were compared using the formula h1-h2 (se = sqrt(se1^2 + se2 ^2)).
Heritability estimates under an additive model were generated using GCTA version 1.26.0 [7 (link)] and PLINK version 1.90b3.32 [8 (link)] by partitioning the genome. To reduce computational time, a smaller control cohort of 10,000 individuals was randomly generated from the UK Biobank and used for each set of cases. SNPs were filtered for deviation from Hardy-Weinberg equilibrium (P > 1 × 10−6) and minor allele frequency >0.01. A genetic relationship matrix was created for each chromosome, which was then used to calculate heritability assuming a prevalence of gout of 2% in the general population.
Cadzow M., Merriman T.R, & Dalbeth N. (2017). Performance of gout definitions for genetic epidemiological studies: analysis of UK Biobank. Arthritis Research & Therapy, 19, 181.
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Publication 2017
Adrenal Cortex Hormones Allopurinol Anti-Inflammatory Agents, Non-Steroidal Arthritis, Gouty Chromosomes Colchicine Diagnosis Ethnicity Europeans Febuxostat Genome Genome-Wide Association Study Gout Kidney Diseases Leukemia Lymphoma Patient Discharge Pharmaceutical Preparations Probenecid Selfish Genes Single Nucleotide Polymorphism Sulfinpyrazone Urate Waist Circumference
4
Quantification of LTB4 in WBC
Cited 8 times
Fresh buffy coat was obtained from hospital blood bank and subjected for WBC separation using HiSep (Hi-media, Mumbai) and diluted with equal volume of Hank’s balanced salt solution. The metabolism of endogenously bound arachidonic acid to LTB4 was measured in a total volume of 0.4 ml of the cell suspension at 37°C. The reaction tubes were containing 20 μg arachidonic acid and the investigational drugs (PHF and zileuton). After 45 min pre-incubation the reaction was started by addition of 20 μg of calcium ionophore and 2 μg of glutathione. After 15 min the reaction was stopped with 40 μl of 0.1 M HCl. After centrifugation for 2 min aliquots of the supernatant were subjected to LC-MS/MS estimation of LTB4.
Hypersil GOLD column (50 mm × 2.1 mm, 1.9 μm, Thermo, Waltham, MA, USA) with a gradient system of acetonitrile with 0.1% formic acid and water with 0.1% formic acid was used for chromatographic separation. The gradient was started with 80:20 water/acetonitrile and reached to 30:70 in 3 min and first line condition was achieved over a period of 2 min. Flow rate was maintained at 0.5 ml/min.
Tandem Mass spectrometric detection of analytes and internal standard (IS) was carried out with an electrospray ionization (ESI) source operated in the negative mode. Multiple Reaction Monitoring (MRM) mode was performed for quantification. 335.2 m/z was selected as precursor ion (Q1) whereas 317.2 m/z and 195.1 m/z (Q3) were selected as product ion for LTB4. Optimized compound dependent parameters were Declustering Potential (DP) - 76, Entrance Potential (EP) - 10, Collision Energy (CE) - 20 and Collision Cell Exit Potential (CXP) - 8. Probenecid was selected as an IS with 283.91 m/z (Q1) and 240 m/z (Q3).
Velpandian T., Gupta P., Ravi A.K., Sharma H.P, & Biswas N.R. (2013). Evaluation of pharmacological activities and assessment of intraocular penetration of an ayurvedic polyherbal eye drop (Itone™) in experimental models. BMC Complementary and Alternative Medicine, 13, 1.
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Publication 2013
acetonitrile Arachidonic Acid Calcium Ionophores Cells Centrifugation Chromatography formic acid Glutathione Gold Investigational New Drugs Leukotriene B4 Metabolism Probenecid Sodium Chloride Tandem Mass Spectrometry zileuton
5
TRPA1 Calcium Influx Assay
Cited 8 times
HEK-293 cells stably expressing human TRPA1 were plated into 384-well plates at a density of 20,000 cells/well 24 hours prior to assaying. On the day of assay, cells were loaded with 4 μM Fluo-4 dye (Invitrogen) and 0.08% pluronic acid (Invitrogen) for 1 hour at room temperature in assay buffer consisting of Hank's balanced salt solution (Invitrogen) supplemented with 20 mM HEPES (Invitrogen), 2.5 mM probenecid (Sigma, St. Louis, MO), and 4% TR-40 (Invitrogen). Calcium influx assays were performed using the Fluorometric Imaging Plate Reader (FLIPR) TETRA (Molecular Devices, Sunnyvale, CA). Concentration-response curves were generated for the TRPA1 agonists cinnamaldehyde and AITC prior to antagonist testing so EC60 concentrations could be determined. Titrations of HC-030031 were made from a DMSO stock solution and DMSO was kept to a constant of 0.4% in the assay. The antagonist was incubated with the cells for 10 minutes before the addition of an EC60 concentration of either cinnamaldehyde (18 μM) or AITC (6 μM) and calcium influx was monitored for an additional 10 minutes.
Eid S.R., Crown E.D., Moore E.L., Liang H.A., Choong K.C., Dima S., Henze D.A., Kane S.A, & Urban M.O. (2008). HC-030031, a TRPA1 selective antagonist, attenuates inflammatory- and neuropathy-induced mechanical hypersensitivity. Molecular Pain, 4, 48.
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Publication 2008
2,3,4-tri-O-acetylarabinopyranosyl isothiocyanate Acids agonists Biological Assay Buffers Calcium Cells cinnamic aldehyde Fluo 4 Fluorometry Hanks Balanced Salt Solution HC 030031 HEK293 Cells HEPES Homo sapiens Medical Devices Pluronics Probenecid Sulfoxide, Dimethyl Tetragonopterus Titrimetry

Most recents protocols related to «Probenecid»

1
Tyrode and Extracellular Solutions for Voltage-Clamp Experiments
Tyrode solution contained (in mM) 140 NaCl, 5 KCl, 2.5 CaCl2, 2 MgCl2, and 10 HEPES. The standard extracellular solution for voltage-clamp experiments contained (in mM) 140 TEA-methane-sulfonate, 2.5 CaCl2, 2 MgCl2, 1 4-aminopyridine, 10 HEPES, and 0.002 tetrodotoxin. For the experiments described in Figs. 1 and 2, the extracellular solution also contained 0.33% DMSO. The standard pipette solution contained (in mM) 120 K-glutamate, 5 Na2-ATP, 5 Na2-phosphocreatine, 5.5 MgCl2, 5 glucose, and 5 HEPES. For measurements of rhod-2 Ca2+ transients, it also contained 15 EGTA, 6 CaCl2, and 0.1 rhod-2. For measurements with fluo-4, isolated muscle fibers were incubated for 30 min in the presence of Tyrode solution containing 10 μM fluo-4 AM. All solutions were adjusted to pH 7.20. The Ringer solution used for muscle force measurements contained (in mM) 140 NaCl, 6 KCl, 3 CaCl2, 2 MgCl2, and 10 HEPES, adjusted to pH 7.40.
Probenecid was prepared as a 0.3 M aliquoted stock solution in DMSO and used in the extracellular solution at 0.5, 1, or 2 mM. Carbenoxolone was prepared as a 10 mM stock solution in the extracellular solution and used at 0.1 mM. These concentrations were chosen on the basis of their effectiveness and wide use to block Panx1 channels throughout the literature (e.g., Dahl et al., 2013 (link)). When testing the effect of either probenecid or carbenoxolone using the preincubation protocol (Figs. 1 and 2), fibers were bathed in the drug-containing extracellular solution from the beginning of the intracellular dialysis with the rhod-2-containing solution (i.e., 30 min before taking measurements). The 10panx1 peptide and the scrambled control peptide (10panx1SCr) were tested under the same conditions at 200 µM while the P2Y2 antagonist AR-C 118925XX was tested at 10 µM. All chemicals and drugs were purchased from Sigma-Aldrich, except for tetrodotoxin (Alomone Labs), rhod-2 and fluo-4 (Thermo Fisher Scientific), and AR-C 118925XX (TOCRIS—Bio-Techne).
In vitro fluorescence measurements using droplets of a solution containing (in mM) 120 K-glutamate, 10 HEPES, 15 EGTA, 6 CaCl2, and 0.1 rhod-2, with or without probenecid, showed that fluorescence intensity in the presence of 1 mM probenecid corresponded to 1.09 ± 0.12% (n = 6) the intensity in the absence of probenecid, excluding an interaction of the drug with the dye to explain the effect on resting fluorescence in muscle fibers.
Jaque-Fernandez F., Allard B., Monteiro L., Lafoux A., Huchet C., Jaimovich E., Berthier C, & Jacquemond V. (2023). Probenecid affects muscle Ca2+ homeostasis and contraction independently from pannexin channel block. The Journal of General Physiology, 155(4), e202213203.
Publication 2023
Aminopyridines Carbenoxolone Cardiac Arrest Dialysis Solutions Drug Interactions Egtazic Acid Figs Fluo 4 Fluorescence Glucose Glutamate HEPES Magnesium Chloride methanesulfonate Muscle Tissue P2RY2 protein, human Peptides Pharmaceutical Preparations Phosphocreatine Probenecid Protoplasm rhod-2 Ringer's Solution Sodium Chloride Sulfoxide, Dimethyl Tetrodotoxin Transients Tyrode's solution
2
Quantifying Voltage-Dependent Ca2+ Release Dynamics
Detection of fluorescence from the Ca2+-sensitive dye dialyzed into the fiber’s cytosol was achieved with a Zeiss LSM 800 microscope equipped with a 63× oil immersion objective (numerical aperture 1.4). Standard green and red configurations were used for detection of the fluorescence of fluo-4 and rhod-2, respectively. Voltage-activated fluorescence changes were imaged using the line-scan mode (x,t) of the system. They were expressed as F/F0 with F0 as the baseline fluorescence. In experiments designed to follow the effect of probenecid after acute application of the drug, the changes in resting fluorescence F0 were normalized to F0 in the control condition at the beginning of the experiment; this value is referred to as intialF0. In graphs presenting fluorescence transients, the y-scale bar corresponds to the indicated multiple of F0. Quantification of the Ca2+ release flux (dCaTot/dt) underlying the rhod-2 Ca2+ transients was performed as previously described (Lefebvre et al., 2011 (link); Kutchukian et al., 2016 (link)). The resting Ca2+ concentration was assumed to be 100 nM in all conditions, except for results shown in Figs. 8 and 9, for which the change in resting Ca2+ concentration induced by probenecid was implemented in the Ca2+ release flux calculation. For this, the resting Ca2+ level in the presence of probenecid was calculated from the increase in resting fluorescence, assuming an initial resting [Ca2+] level of 0.1 µM, and Fmax/Fmin and KD values for rhod-2 of 30 and 1.63 µM, respectively (Sanchez et al., 2021 (link)). The voltage-dependence of the peak Ca2+ release flux from each muscle fiber was fitted with a Boltzmann function: dCaTotdt=MaxdCaTotdt1+expV0.5−V/k, with Max dCaTot/dt the maximum Ca2+ release flux, V0.5 the mid-activation voltage, and k the steepness factor.
Jaque-Fernandez F., Allard B., Monteiro L., Lafoux A., Huchet C., Jaimovich E., Berthier C, & Jacquemond V. (2023). Probenecid affects muscle Ca2+ homeostasis and contraction independently from pannexin channel block. The Journal of General Physiology, 155(4), e202213203.
Publication 2023
Cytosol Figs Fluo 4 Fluorescence Microscopy Muscle Tissue Pharmaceutical Preparations Probenecid Radionuclide Imaging rhod-2 Submersion Transients
3
Probenecid Effects on Isolated Muscle Force
Electrically stimulated isolated extensor digitorum longus (EDL) muscles were used to assess the effect of probenecid on force development. For this, 8-wk-old OF1 male mice were anesthetized by isoflurane inhalation and killed by cervical dislocation, and EDL muscles were removed. Force measurements were performed using the 1205A Isolated Muscle System from Aurora Scientific. The EDL muscle was mounted in the experimental chamber in the presence of air-bubbled Ringer solution (see Solutions) maintained at 25°C. The muscle was stretched to the optimum length for the production of maximum isometric twitch force triggered by a supra-maximum 0.2-ms-long electrical stimulation. From this time point, the muscle was repeatedly stimulated by a series of five such electrical stimulations applied at 0.1 Hz, every 5 min, throughout the experiment. Force measurements were performed in response to 60 Hz supramaximum tetanic trains of stimulations of 0.5 s total duration, applied every 15 min. Following the first tetanic response, the Ringer solution was exchanged for the DMSO-containing Ringer with or without probenecid. Force values were normalized by the weight of the muscle.
Jaque-Fernandez F., Allard B., Monteiro L., Lafoux A., Huchet C., Jaimovich E., Berthier C, & Jacquemond V. (2023). Probenecid affects muscle Ca2+ homeostasis and contraction independently from pannexin channel block. The Journal of General Physiology, 155(4), e202213203.
Publication 2023
Electricity Inhalation Isoflurane Joint Dislocations Males Mus Muscle Tissue Neck Probenecid Ringer's Solution Stimulations, Electric Sulfoxide, Dimethyl Toxoid, Tetanus
4
Muscle Fiber Pharmacological Analysis Protocols
During the experiments, muscle fibers and muscles were randomly assigned to the control and test (probenecid, carbenoxolone, or other treatment) groups. When comparing results between control muscle fibers and muscle fibers treated under a given pharmacological condition, groups of fibers from the same animal were considered as technical replicates and statistical comparison was achieved using a nested analysis, as described by Eisner (2021) (link). When determining the extent of change in a given parameter produced by the acute application of a pharmacological compound (with the initial measurement from the same muscle or muscle fiber being the control value), each experiment was considered independent, and statistics were based on the number of fibers.
The number of necessary experiments was estimated on the basis of our previous experience with each protocol. A sample size estimation (with α = 0.05 and β = 0.80) was made from our pilot data showing a 55% reduction of maximum SR Ca2+ release in the presence of 1 mM probenecid, giving a total of six per group.
Experiments on single muscle fibers and isolated muscles require a great extent of specific expertise, preparation, dexterity, and watchfulness, with the operator on duty being in charge from beginning to end. Under these conditions, further complexification by blinding was not implemented to prevent any related risk of misidentification error. Blinding would also have required two dedicated persons on duty on each experimental day, which was not routinely possible.
Data and statistical analysis complied with the recommendations on experimental design and analysis in pharmacology (Curtis et al., 2018 (link)). Electrophysiological and fluorescence data were processed with Clampfit 10.0 (Molecular Devices) and ImageJ software (National Institutes of Health), respectively, and Origin 7.0 (OriginLab Corporation). Statistical analysis was performed with GraphPad Prism 9.1 (GraphPad Software). Data values are presented as means ± SD for either n muscle fibers or n muscles. Individual datapoints for the parameters measured from each and every muscle/fiber tested are also presented in the figures. Statistical comparison was made only on measurements conducted under a given experimental condition on a minimum group of either five distinct muscles or five distinct muscle fibers. No search was made for outliers within datasets. The level of probability (p) deemed to constitute the threshold for statistical significance was defined as P < 0.05. Exact P values are indicated on the graphs in the figures.
For single muscle fiber experiments designed to test the effect of chronic exposure to a pharmacological compound (Figs. 1, 2, and 4), the normality of the distribution of each Boltzmann fit parameter was assessed using Shapiro-Wilk test, and the statistical difference between the control group and either the probenecid group or the carbenoxolone group was determined using the hierarchical (nested) analysis or linear mixed modeling described by Eisner (2021) (link), taking into account the number of fibers from each mouse.
For single muscle fiber experiments designed to test the effect of acute exposure to probenecid, statistical differences between the control group and the probenecid group in terms of the extent of change in (1) baseline rhod-2 fluorescence, (2) resting membrane conductance, and (3) peak SR Ca2+ release flux during the test period and after wash-out, was assessed using unpaired Mann–Whitney Wilcoxon test (Figs. 6 and 9).
For muscle force measurements in control conditions and in the presence of probenecid, a Mann–Whitney Wilcoxon test was used to compare the amplitude of tetanic force at a given time point of the experiment. Friedman’s nonparametric test for repeated measurements followed by Dunn’s post-hoc test was used to compare the time-dependent changes in the tetanic force amplitude in the two groups.
Jaque-Fernandez F., Allard B., Monteiro L., Lafoux A., Huchet C., Jaimovich E., Berthier C, & Jacquemond V. (2023). Probenecid affects muscle Ca2+ homeostasis and contraction independently from pannexin channel block. The Journal of General Physiology, 155(4), e202213203.
Publication 2023
Animals Carbenoxolone Fibrosis Figs Fluorescence Medical Devices Mice, House Muscle Tissue Myopathy prisma Probenecid rhod-2 Tetanus Tissue, Membrane
5
Calcium Mobilization Assay for Opioid Receptors
Stock solutions (10 mM) of the peptides were prepared in 5% DMSO in bidistilled water and kept at −20°C Serial dilutions were carried out in HBSS/HEPES buffer (20 mM, containing 0.02% bovine serum albumin fraction V). Calcium mobilization assay was performed using the same method as previously described (Camarda and Calo, 2013 (link)). CHO cells with stable co-expression of human mu or kappa receptors and the C-terminally modified Gαqi5 and CHO cells with co-expression of the delta-opioid receptor and the GαqG66Di5 protein were used. Dulbecco’s MEM/HAMS F12 (1:1) medium supplemented with 10% fetal bovine serum, penicillin (100 IU/mL), streptomycin (100 μg/mL), L-glutammine (2 mM), fungizone (1 μg/mL), geneticin (G418; 200 μg/mL) and hygromycin B (100 μg/mL) was used for cell culture. Cells were seeded at a density of 50,000 cells/well into 96-well black, clear-bottom plates and kept in the incubator at 37°C in 5% CO2/humidified air. After 24 h the cell growth medium was aspired and loading medium, supplemented with probenecid (2.5 mM), calcium sensitive fluorescent dye Fluo-4 AM (3 µM), pluronic acid (0.01%) and HEPES (20 mM), was added. Then the plates were placed in the incubator again. After 30 min the loading solution was aspirated and 100 µL/well of assay buffer (HBSS supplemented with 20 mM HEPES, 2.5 mM probenecid, and 500 µM Brilliant Black) was added. Next, both plates (cell culture and compound plate) were placed in the FlexStation II reader (Molecular Device, Union City, CA 94587, United States), the on-line additions were carried out in a volume of 50 µL/well and the fluorescence changes were measured. Ligand efficacies, expressed as the intrinsic activity (α), were calculated as the Emax ratio of the tested compound and the standard agonist. At least three independent experiments for each assay were carried out in duplicate.
Curve fittings were performed using Graph Pad PRISM 5.0 (GraphPad Software Inc., San Diego, United States). Data have been statistically analyzed with one way ANOVA followed by the Dunnett’s test for multiple comparisons; p values < 0.05 were considered significant.
Piekielna-Ciesielska J., Malfacini D., Djeujo F.M., Marconato C., Wtorek K., Calo’ G, & Janecka A. (2023). Functional selectivity of EM-2 analogs at the mu-opioid receptor. Frontiers in Pharmacology, 14, 1133961.
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Publication 2023
Acids antibiotic G 418 ATF7IP protein, human Biological Assay brilliant black Buffers Calcium Cell Culture Techniques Cells CHO Cells Fetal Bovine Serum Fluo 4 Fluorescence Fluorescent Dyes Fungizone Geneticin Hemoglobin, Sickle HEPES Homo sapiens Hygromycin B Ligands Medical Devices neuro-oncological ventral antigen 2, human Penicillins Peptides Pluronics prisma Probenecid Proteins Receptors, Cell Surface Receptors, Opioid, delta Receptors, Opioid, kappa Receptors, Opioid, mu Serial C Serum Albumin, Bovine Streptomycin Sulfoxide, Dimethyl Technique, Dilution

Top products related to «Probenecid»

1
Probenecid by Merck Group
Sourced in United States, Germany, France, Belgium, Sao Tome and Principe, Japan, Italy, Canada, United Kingdom, Switzerland, Spain, Macao, Austria
Probenecid is a laboratory reagent used in the analysis of biological samples. It functions as a uricosuric agent, which helps to increase the excretion of uric acid from the body. Probenecid is commonly used in various biochemical and clinical applications, such as drug interaction studies and research on uric acid metabolism.
2
Probenecid by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Canada
Probenecid is a pharmaceutical compound used to block the reabsorption of uric acid in the kidney. It is primarily used as an adjunct therapy in the treatment of gout and hyperuricemia. Probenecid functions by inhibiting the organic anion transporters responsible for the reabsorption of uric acid, leading to increased urinary excretion of uric acid.
3
Fluo 4 am by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, Canada, China, France, Spain, Italy, Sweden, Japan, Switzerland, Belgium, Australia, Austria, Finland, New Zealand
Fluo-4 AM is a fluorescent calcium indicator used for the detection and measurement of intracellular calcium levels. It functions by binding to calcium ions, which results in an increase in fluorescence intensity. This product is commonly used in various cell-based assays and research applications involving calcium signaling.
4
Flexstation 3 by Molecular Devices
Sourced in United States, United Kingdom, Japan, Germany, Switzerland, Spain, China
The FlexStation 3 is a multimode microplate reader that measures various assays, including fluorescence, luminescence, and absorbance. It is designed to provide consistent and reliable results for a wide range of applications in life science research and drug discovery.
5
Pluronic f 127 by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom, Japan, Belgium, Italy, Lithuania, Canada, India
Pluronic F-127 is a non-ionic, surfactant-based material commonly used in various laboratory applications. It is a triblock copolymer composed of polyethylene oxide and polypropylene oxide segments. Pluronic F-127 is known for its ability to form thermoreversible gels and has versatile applications in areas such as drug delivery, tissue engineering, and cell culture.
6
Flipr tetra by Molecular Devices
Sourced in United States, United Kingdom
The FLIPR Tetra is a high-throughput cellular screening system designed for rapid and sensitive detection of cellular responses. It utilizes fluorescence-based detection to measure changes in cellular parameters, such as membrane potential and calcium flux, in real-time. The FLIPR Tetra is capable of simultaneously monitoring multiple wells in a microplate format, making it a versatile tool for a wide range of applications in drug discovery and cell biology research.
7
Fura 2 am by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, Italy, France, Spain, Australia, Japan, China, Canada, Belgium, Austria, Hungary, Macao
Fura-2 AM is a fluorescent calcium indicator used for measuring intracellular calcium levels. It is a cell-permeable derivative of the parent compound Fura-2. Fura-2 AM can be loaded into cells, where intracellular esterases cleave off the acetoxymethyl (AM) ester group, trapping the Fura-2 indicator inside the cell.
8
Fluo 4 nw calcium assay kit by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Italy
The Fluo-4 NW calcium assay kit is a fluorescence-based tool used to measure intracellular calcium levels in living cells. It utilizes the Fluo-4 NW fluorescent dye, which binds to calcium ions and emits a fluorescent signal upon excitation. The kit provides a quick and reliable method for detecting and monitoring changes in calcium concentrations within cellular environments.
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Hepes by Merck Group
Sourced in United States, Germany, United Kingdom, France, Canada, Switzerland, Italy, China, Belgium, Spain, Sao Tome and Principe, Australia, Japan, India, Macao, Brazil, Ireland, Denmark, Austria, Poland, Netherlands, Sweden, Norway, Israel, Hungary, Holy See (Vatican City State), Mexico
HEPES is a buffering agent commonly used in cell culture and biochemical applications. It helps maintain a stable pH environment for biological processes.
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Bovine serum albumin (bsa) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Japan, France, Sao Tome and Principe, Canada, Macao, Spain, Switzerland, Australia, India, Israel, Belgium, Poland, Sweden, Denmark, Ireland, Hungary, Netherlands, Czechia, Brazil, Austria, Singapore, Portugal, Panama, Chile, Senegal, Morocco, Slovenia, New Zealand, Finland, Thailand, Uruguay, Argentina, Saudi Arabia, Romania, Greece, Mexico
Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.

More about "Probenecid"

Probenecid is a uricosuric agent commonly used to treat gout and hyperuricemia, a condition characterized by high levels of uric acid in the blood.
It works by inhibiting the reabsorption of uric acid in the kidneys, leading to increased urinary excretion.
This medication may also be used to prevent or treat certain types of kidney stones.
Probenecid is often prescribed in combination with other drugs, such as antibiotics, to enhance their effectiveness.
While generally well-tolerated, probenecid may cause side effects like gastrointestinal discomfort, rash, and headache in some patients.
Monitoring of uric acid levels and kidney function is recommended during treatment.
Probenecid is an important tool in the management of gout and related conditions, offering an effective way to lower uric acid levels and improve patient outcomes.
Other related compounds and terms include Fluo-4 AM, a fluorescent calcium indicator used in calcium assays, FlexStation 3, a microplate reader used for these assays, Pluronic F-127, a surfactant used to solubilize Fluo-4 AM, FLIPR Tetra, a high-throughput screening system for calcium-based assays, Fura-2 AM, another fluorescent calcium indicator, and the Fluo-4 NW calcium assay kit.
HEPES and bovine serum albumin (BSA) are also commonly used in these types of calcium-based experiments and assays.
By understanding the broader context and related terms, researchers can optimize their probenecid-related studies and improve the reproducibility and accuracy of their findings.