Mk571

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, Sao Tome and Principe, Macao

MK571 is a laboratory equipment product manufactured by Merck Group. It is a multi-function device designed for various scientific and research applications. The core function of MK571 is to provide precise measurements and analyses for researchers and scientists.

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Lab products found in correlation

Ko143, by Merck Group (22 mentions) Verapamil, by Merck Group (12 mentions) Rhodamine 123, by Merck Group (10 mentions) Ripa buffer, by Thermo Fisher Scientific (9 mentions) Dimethyl sulfoxide (dmso), by Merck Group (8 mentions) Fumitremorgin c, by Merck Group (7 mentions) Doxorubicin, by Merck Group (6 mentions) Vinblastine, by Merck Group (5 mentions) Psc833, by Merck Group (5 mentions) Alamethicin, by Merck Group (4 mentions) Cyclosporin a, by Merck Group (4 mentions) Fetal bovine serum (fbs), by Merck Group (4 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (4 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (4 mentions) Mitoxantrone, by Merck Group (3 mentions) Calcein am, by Merck Group (3 mentions) Forskolin, by Merck Group (3 mentions) Cm dcfda, by Merck Group (3 mentions) Fl bopidy, by Merck Group (3 mentions)

74 protocols using mk571

Microfluidic Fluorescence Imaging Assay

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Medium in all perfusion channels was replaced with 1.25 μM CMFDA (Molecular Probes, C7025, stock 2.5 mM in DMSO) in the presence of 0, 10, 20, and 30 μM MK571 (Sigma, M7571, stock 10 mM in H 2 O) in KHH buffer. After a 30-min incubation on the rocker platform, the chips were washed one time with ice cold KHH buffer. In the next washing step, Hoechst 33342 (2 drops/mL, Life Technologies, R37605), 10 μM PSC833 (Sigma, stock 5 mM in DMSO), 10 μM Ko143 (Sigma, K2144, stock 10 mM in DMSO), and 10 μM MK571 (Sigma, M7571) were added to the washing solution and the plate was imaged with the ImageXpress® Micro Confocal High-Content Imaging System.

Vormann M.K., Gijzen L., Hutter S., Boot L., Nicolas A., van den Heuvel A., Vriend J., Ng C.P., Nieskens T.T.G., van Duinen V., de Wagenaar B., Masereeuw R., Suter-Dick L., Trietsch S.J., Wilmer M., Joore J., Vulto P, & Lanz H.L. (2018). Nephrotoxicity and Kidney Transport Assessment on 3D Perfused Proximal Tubules. The AAPS journal, 20(5).

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Oxidative Stress Enzyme Assay

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Benznidazole (BZL), dithionitrobenzoic acid, oxidized glutathione (GSSH), reduced glutathione (GSH), glutathione, nitroblue tetrazolium, MK571, β-NADPH, riboflavin, rifampicin (RIF), sulfosalicylic acid, tert-butyl hydroperoxide (tBOOH) and 2-vinylpyridine were purchased from Sigma-Aldrich (St. Louis, MO, USA). DMSO and hydrogen peroxide were purchased from Merck (Darmstadt, HE, Germany). All other chemicals were of analytical grade purity.

Rigalli J.P., Perdomo V.G., Ciriaci N., Francés D.E., Ronco M.T., Bataille A.M., Ghanem C.I., Ruiz M.L., Manautou J.E, & Catania V.A. (2016). The antitripanocide Benznidazole promotes adaptive response to oxidative injury: Involvement of the nuclear factor-erythroid 2-related factor-2 (Nrf2) and multidrug resistance associated protein 2 (MRP2). Toxicology and applied pharmacology, 304, 90-98.

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Quantitative Analysis of Lipophilic Drug Transport

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Dimethyl sulfoxide (DMSO), insulin, 1,6-diphenyl-1,3,5-hexatriene (DPH), stearic acid, oleic acid, tamoxifen, valspodar (PSC833), calcein-am, sulforhodamine 101, Ko143, MK571, and mitoxantrone were purchased from Sigma-Aldrich (St Louis, MO, USA). Hydrocortisone hemisuccinate was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Penicillin/streptomycin, fetal bovine serum (FBS) were purchased from Biochrom (Berlin, Germany). 7-β-NBD-taurocholate was custom-synthesized as described by Schneider et al. [34 (link)] PPC and PI were obtained from Lipoid (Ludwigshafen am Rhein, Germany). EPL (Essentiale Forte 300 mg) was obtained from Sanofi. All other chemicals were purchased from commercial sources and were of the highest purity available.

Wupperfeld D., Fricker G., Bois De Fer B., Frank L., Wehrle A, & Popovic B. (2022). Essential phospholipids decrease apoptosis and increase membrane transport in human hepatocyte cell lines. Lipids in Health and Disease, 21, 91.

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Glioblastoma Cell Growth Inhibition

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Glioblastoma cells U251, MZ-256, and MZ-327 were pre-treated with the MRP1 small molecule inhibitor MK571 (M7571 SIGMA) and Reversan (SML0173 SIGMA) at a final concentration of 25 and 15 μM; respectively, for 7 h and subsequently were seeded at 1 × 10⁴ cells/well in a 6 well plate and allowed to adhere overnight. Cells were then allowed to grow for 96 h and counted using the trypan blue exclusion method.

Tivnan A., Zakaria Z., O'Leary C., Kögel D., Pokorny J.L., Sarkaria J.N, & Prehn J.H. (2015). Inhibition of multidrug resistance protein 1 (MRP1) improves chemotherapy drug response in primary and recurrent glioblastoma multiforme. Frontiers in Neuroscience, 9, 218.

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Prostanoid and Nucleotide Quantification

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All prostanoids (PGE₃, PGF_3α, TXB₃, PGE₂, PGF_2α, and TXB₂), deuterated prostanoids (PGE₂-d4, PGF_2α-d4, and TXB₂-d4), and EPA were purchased from Cayman Chemical Co. (Ann Arbor, MI). Calcium ionophore A23187 and indomethacin were purchased from Sigma Aldrich (St. Louis, MO). Adenosine 5′-triphosphate (ATP) disodium salt hydrate, 5′-adenylic acid (AMP), and sodium creatine phosphate hydrate were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Creatine kinase was purchased from Roche Applied Science (Tokyo, Japan). Dipyridamole, probenecid, quercetin, methotrexate, and folic acid were purchased from Wako Pure Chemical Industries (Osaka, Japan). MK571, celecoxib, candesartan, estradiol 17β-glucronide (E₂17βG), adenosine 3′, 5′-cyclic monophosphate (cAMP), and guanosine 3′, 5′-cyclic monophosphate (cGMP) were purchased from Sigma Aldrich (St. Louis, MO). All other chemicals were of the highest purity available.

Tanaka N., Yamaguchi H, & Mano N. (2014). Transport of Eicosapentaenoic Acid-Derived PGE3, PGF3α, and TXB3 by ABCC4. PLoS ONE, 9(10), e109270.

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Preparation and Storage of Enterotoxins

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Synthetic heat-stable enterotoxin (BA Chem, Bubendorf, Switzerland) was resuspended in 100 mM acetic acid, 0.1% BSA, and frozen in single-use aliquots. LT and LT-E112K were reconstituted in water and stored at 4°C. IBMX, MK-571, vardenafil, forskolin, and buffer components were acquired from Sigma (St. Louis, Missouri). Tissue culture media was obtained from ThermoFisher Scientific (Waltham, Massachusetts)

Foulke-Abel J., Yu H., Sunuwar L., Lin R., Fleckenstein J.M., Kaper J.B, & Donowitz M. (2020). Phosphodiesterase 5 (PDE5) restricts intracellular cGMP accumulation during enterotoxigenic Escherichia coli infection. Gut Microbes, 12(1), 1752125.

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Astroglial Urate Neuroprotection Assay

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MK-571 and BSO were purchased from Sigma and dissolved in DMEM as 100× concentrated stocks. Enriched astroglial cultures were treated with vehicle or urate (100 μM) alone or in combination with BSO (0.25 mM) or MK-571 (50 μM). Twenty-four hours later CM was collected and filtered through a 0.2 μm membrane to remove cellular debris and immediately used for experiments with MES 23.5 cells. To verify the effects of BSO or MK-571, GSH content in the CM was measured. MES 23.5 cells were pretreated with vehicle CM or urate CM, with or without BSO or MK-571, (or were not pretreated) before being exposed to oxidative stressor (H₂O₂) for 24h. The percentage of dead cells was analyzed using the FACS method described above.

Bakshi R., Zhang H., Logan R., Joshi I., Xu Y., Chen X, & Schwarzschild M.A. (2015). Neuroprotective effects of urate are mediated by augmenting astrocytic glutathione synthesis and release. Neurobiology of disease, 82, 574-579.

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Quantifying Efflux Pump Activity in Cells

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Cells were incubated in the presence of 10 µM Rhodamine 123 (P-gp substrate, Sigma), FL-BOPIDY (BCRP substrate, Sigma), or CM-DCFDA (MRP substrate, Sigma) for 1 h at 37 ºC followed by cell lysis using RIPA buffer (ThermoFisher). For assessing the contribution of efflux pump in the drug uptake, cells were pre-incubated for 1 h in presence of 5 µM cyclosporine A (CsA, P-gp inhibitor, Sigma), 1 µM Ko143 (BCRP inhibitor, Sigma), or 10 µM MK571 (MRPs inhibitor, Sigma) and maintained during the incubation with drug efflux substrate. Following incubation, cells were briefly washed with ice-cold PBS and lysed with RIPA buffer. Fluorescence in cell lysates was assessed using a SynergyMX² ELISA plate reader (Bio-Tek, Winooski, VT, USA). Relative fluorescence units (RFU) were normalized against the total protein content and the protein levels were determined by bicinchoninic acid assay (BCA, ThermoFisher). Fluorescence values (expressed as relative fluorescence unit or RFU) obtained from cell lysates in the absence of inhibitor (named as controls) were normalized to the protein content and expressed as RFU/µg protein.

Raut S., Patel R, & Al-Ahmad A.J. (2021). Presence of a mutation in PSEN1 or PSEN2 gene is associated with an impaired brain endothelial cell phenotype in vitro. Fluids and Barriers of the CNS, 18, 3.

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Assessing BBB Transporter Function

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P-gp, MRP, and BCRP functionality were assessed using rhodamine 123 (Sigma), DiOC2 (3,3′-diethyloxacarbocyanine Iodide) (Sigma), citalopram (Sigma), and doxorubicin (Sigma). Both channels were pretreated with 50 µM verapamil (Sigma), 10 µM MK571 (Sigma), or 1 µM Ko143, which are inhibitors of P-gp, MRPs, or BCRP, respectively. At 30 min after pre-treatment with inhibitors, rhodamine 123 (2 µM), DiOC2 (2 µM), citalopram or doxorubicin (5 µM) in the presence or absence of inhibitor was dosed to brain endothelial channel with flow rate at 100 µl h⁻¹. To monitor the barrier integrity, 100 µg ml⁻¹ of 3 kDa dextran-cascade blue (Thermo Fisher) was dosed simultaneously. We collected apical and basal effluent for 6 h and tracers in the effluents were quantified by measuring fluorescent intensity. The fluorescence was measured at 485/530 nm, 482/497 nm, or 470/585 nm to quantify rhodamine 123, DiOC2, or doxorubicin, respectively, using a Synergy H1 microplate reader (BioTek, USA). Amount of citalopram in the apical and basal media was quantified using mass spectroscopy. The increase of BBB permeability of the drugs in the presence of inhibitor was presented as ‘ratio of P_app’.

Park T.E., Mustafaoglu N., Herland A., Hasselkus R., Mannix R., FitzGerald E.A., Prantil-Baun R., Watters A., Henry O., Benz M., Sanchez H., McCrea H.J., Goumnerova L.C., Song H.W., Palecek S.P., Shusta E, & Ingber D.E. (2019). Hypoxia-enhanced Blood-Brain Barrier Chip recapitulates human barrier function and shuttling of drugs and antibodies. Nature Communications, 10, 2621.

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Glucuronidation Profiling of Flavonoids

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Alamethicin, β-estradiol, dipyridamole (DIPY), D-saccharic-1, 4-lactone, magnesium chloride (MgCl₂), MK-571, Ko143, leukotriene C4 (LTC4) and uridine diphosphate glucuronic acid (UDPGA) were all obtained from Sigma-Aldrich (St Louis, MO). Apigenin, chrysin, cycloicaritin, genistein, and wushanicaritin with purity over 98% was purchased from Aladdin Reagents (Shanghai, China). Apigenin-7-O-glucuronide, chrysin-7-O-glucuronide, genistein-7-O-glucuronide, and β-estradiol-3-O-glucuronide was purchased from Toronto Research Chemicals (North York, ON, Canada). Wushanicaritin-3-O-glucuronide was prepared in our laboratory as described previously [23 (link)]. 293T cells, HeLa cells, pLVX-mCMV-ZsGreen-PGK-Puro vector [9371 base pairs (bp)] and pLVX-shRNA2-Neo vector (9070 bp) were all provided from BioWit Technologies (Shenzhen, China). The pGEM-T plasmid which carried the UGT1A1 cDNA clone was purchased from Sino Biological Inc. (Beijing, China), while recombinant human UGT1A1 was purchased from Corning Biosciences (New York, USA). The anti-BCRP, anti-GAPDH, anti-MRP1, anti-MRP2, anti-MRP3, anti-MRP4 and anti-UGT1A1 antibodies were all obtained from OriGene Technologies (Rockville, MD). All other chemicals and reagents were the highest grade commercially available.

Li S., Xu J., Yao Z., Hu L., Qin Z., Gao H., Krausz K.W., Gonzalez F.J, & Yao X. (2018). The roles of breast cancer resistance protein (BCRP/ABCG2) and multidrug resistance-associated proteins (MRPs/ABCCs) in the excretion of cycloicaritin-3-O-glucoronide in UGT1A1-overexpressing HeLa cells. Chemico-biological interactions, 296, 45-56.

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