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Mk571

Manufactured by Bio-Techne
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MK571 is a laboratory equipment product manufactured by Bio-Techne. It serves as a general-purpose instrument for various applications in research and development settings.

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Lab products found in correlation

N acetyl l cysteine, by Merck Group (2 mentions) α tocopherol, by Merck Group (2 mentions) Nsc23766, by Merck Group (2 mentions) Ferrostatin 1, by Merck Group (2 mentions) Quercetin, by Bio-Techne (2 mentions) Jzl184, by Cayman Chemical (2 mentions) Ml239, by Merck Group (2 mentions) Sc 26196, by Bio-Techne (2 mentions) Cp100356, by Merck Group (2 mentions) Obatoclax mesylate, by Selleck Chemicals (2 mentions) Ym155, by Selleck Chemicals (2 mentions) Elacridar, by Merck Group (2 mentions) Ml210, by Merck Group (2 mentions) Sr 2640, by Bio-Techne (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) N acetylcysteine, by Merck Group (1 mentions) Irinotecan, by Bio-Techne (1 mentions) Sq22536, by Bio-Techne (1 mentions) Triton x 100, by Merck Group (1 mentions) D glucose, by Merck Group (1 mentions)

8 protocols using mk571

1

Synthesis of Pranlukast Analogues

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A similar preparation to that by Raposo et al. (1996) and Green et al. (Geen et al., 2006 ) was followed for synthesis of pranlukast analogues. A detailed description of compound synthesis and compound characterisation can be found in the associated supporting information section. Pranlukast, zafirlukast and montelukast were purchased from AK-Scientific. Cinalukast, FPL 55712, SR 2640, and MK 571 were purchased from Tocris.
Sleebs B.E., Jarman K.E., Frolich S., Wong W., Healer J., Dai W., Lucet I.S., Wilson D.W, & Cowman A.F. (2020). Development and application of a high-throughput screening assay for identification of small molecule inhibitors of the P. falciparum reticulocyte binding-like homologue 5 protein. International Journal for Parasitology: Drugs and Drug Resistance, 14, 188-200.
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2

Investigating Adenylyl Cyclase Regulation

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N-acetyl cysteine and ATP were purchased from Sigma, Glutathione monoethyl ester from Calbiochem, KH7 and SQ22536, forskolin, Inhibitor-172, MK571 and irinotecan were purchased from Tocris. [14C] 6-Mercaptopurine (6-MP) (51 mCi/mmol) was derived from Moravek Biochemicals, CA. ADCY1, 3,6,7,9 and10 adenylyl cyclase antibodies were purchased from Abcam.
Schnúr A., Premchandar A., Bagdany M, & Lukacs G.L. (2019). Phosphorylation-dependent modulation of CFTR macromolecular signalling complex activity by cigarette smoke condensate in airway epithelia. Scientific Reports, 9, 12706.
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3

Fluorescence Imaging of ABC Transporters

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NBD-CSA was custom synthesized [3 (link), 8 (link), 12 (link)]. Bodipy-Prazosin was purchased from Life Technologies (Grand Island, NY). Sulforhodamine 101 free acid (Texas Red), 4 % paraformaldehyde, 0.2% glutaraldehyde, Triton X-100 and D-glucose were purchased from Sigma-Aldrich (St. Louis, MO). Valspodar (PSC-833), KO143 and MK571 were purchased from Tocris Bioscience (Ellisville, MO). All the reagents used in this study were analytical or best available pharmaceutical grade.
Chan G.N., Evans R., Banks D., Mesev E., Miller D.S, & Cannon R.E. (2016). Selective Induction of P-glycoprotein at the CNS Barriers during Symptomatic Stage of an ALS Animal Model. Neuroscience letters, 639, 103-113.
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4

Small Molecule Acquisition and Preparation

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JZL184 (Cat. no. 13158; ≥97% purity) was purchased from Cayman Chemical. ML239 (SML0442; ≥98%), NSC23766 (SML0952; ≥97%), CP-100356 (PZ0171; ≥98%), elacridar (SML0486; ≥98%), ML210 (SML0521; ≥98%), ferrostatin-1 (SML0583; ≥95%), α-tocopherol (T3251; ≥99%), and N-acetyl-L-cysteine (A7250; ≥99%) were purchased from Sigma. YM-155 (S1130; ≥99%), obatoclax mesylate (S1057; ≥99%), and RITA (S2781; 99%) were purchased from SelleckChem. SC-26196 (4189; 99.2%), quercetin (1125; >98%), and MK-571 (2238; >96.9%) were purchased from Tocris. Austocystin D was purchased from eMolecules. The selective CYP2J2 inhibitor 1-(4-bromophenyl)-4-[4-(hydroxydiphenylmethyl)piperidin-1-yl]butan-1-one was synthesized in-house according to the published methodology (>95% purity)48 (link). All small molecules were dissolved in DMSO, except N-acetylcysteine, which was dissolved in cell culture medium then adjusted to pH 7.5.
Rees M.G., Seashore-Ludlow B., Cheah J.H., Adams D.J., Price E.V., Gill S., Javaid S., Coletti M.E., Jones V.L., Bodycombe N.E., Soule C.K., Alexander B., Li A., Montgomery P., Kotz J.D., Hon C.S., Munoz B., Liefeld T., Dančík V., Haber D.A., Clish C.B., Bittker J.A., Palmer M., Wagner B.K., Clemons P.A., Shamji A.F, & Schreiber S.L. (2015). Correlating chemical sensitivity and basal gene expression reveals mechanism of action. Nature chemical biology, 12(2), 109-116.
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5

Functional Assays for MRP1 and BCRP

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Calcein AM (ThermoFisher, NY, USA) cellular accumulation assays were used for MRP1 function, while BCRP function was assessed with Hoechst 33342 (ThermoFisher, NY, USA). MK571 (Tocris, Bristol, UK) and KO143 (Tocris, Bristol, UK), specific MRP1 and BCRP inhibitors, respectively, were used in the functional assay. M1, M2, and unstimulated U937 cells were washed and resuspended in serum-free RPMI, and then seeded in 96-well Black Clear-Bottom Plates (Costar, Washington, DC, USA). Plates were incubated at 37°C with or without inhibitor (MK571, 10 min incubation; KO143, 2 h incubation). After incubation, 10 μM Calcein AM or 10 μM Hoechst 33342 was added to the plate. Plates were immediately placed in an FLx800 Fluorescence Reader (BioTek, Winooski, VT, USA) for 60 min, and read at 485/528 (ex/em). Cell viability was determined via trypan blue staining.
He H., Buckley M., Britton B., Mu Y., Warner K., Kumar S, & Cory T.J. (2018). Polarized macrophage subsets differentially express the drug efflux transporters MRP1 and BCRP, resulting in altered HIV production. Antiviral Chemistry & Chemotherapy, 26, 2040206617745168.
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6

Small Molecule Acquisition and Preparation

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JZL184 (Cat. no. 13158; ≥97% purity) was purchased from Cayman Chemical. ML239 (SML0442; ≥98%), NSC23766 (SML0952; ≥97%), CP-100356 (PZ0171; ≥98%), elacridar (SML0486; ≥98%), ML210 (SML0521; ≥98%), ferrostatin-1 (SML0583; ≥95%), α-tocopherol (T3251; ≥99%), and N-acetyl-L-cysteine (A7250; ≥99%) were purchased from Sigma. YM-155 (S1130; ≥99%), obatoclax mesylate (S1057; ≥99%), and RITA (S2781; 99%) were purchased from SelleckChem. SC-26196 (4189; 99.2%), quercetin (1125; >98%), and MK-571 (2238; >96.9%) were purchased from Tocris. Austocystin D was purchased from eMolecules. The selective CYP2J2 inhibitor 1-(4-bromophenyl)-4-[4-(hydroxydiphenylmethyl)piperidin-1-yl]butan-1-one was synthesized in-house according to the published methodology (>95% purity)48 (link). All small molecules were dissolved in DMSO, except N-acetylcysteine, which was dissolved in cell culture medium then adjusted to pH 7.5.
Rees M.G., Seashore-Ludlow B., Cheah J.H., Adams D.J., Price E.V., Gill S., Javaid S., Coletti M.E., Jones V.L., Bodycombe N.E., Soule C.K., Alexander B., Li A., Montgomery P., Kotz J.D., Hon C.S., Munoz B., Liefeld T., Dančík V., Haber D.A., Clish C.B., Bittker J.A., Palmer M., Wagner B.K., Clemons P.A., Shamji A.F, & Schreiber S.L. (2015). Correlating chemical sensitivity and basal gene expression reveals mechanism of action. Nature chemical biology, 12(2), 109-116.
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7

Compound Screening for Ion Channel Modulators

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All materials were obtained from Sigma-Aldrich® (St. Louis, MO) unless otherwise specified. Tetrodotoxin was purchased from Calbiochem, EMD Biosciences (La Jolla, CA). MK571, T16Ainh-A01, Eact and albuterol were purchased from Tocris (Minneapolis, MN). Membrane potential dye [fluorescent imaging plate reader (FLIPR) blue reagent] was obtained from Molecular Devices (Sunnyvale, CA). Fura-2, AM and mag-fluo-4, AM were obtained from Molecular Probes. B25 (5-[(2,6-difluorobenzyl)oxy]-2-(2-naphthyl)benzofuran-3-carboxylic acid) was synthesized as in Kumar et al.17 (link) MONNA (N-((4-methoxy)-2-naphthyl)-5-nitroanthranilic acid) was a gift from Dr. C. Justin Lee from Korea Institute of Science and Technology.
Danielsson J., Perez-Zoghbi J., Bernstein K., Barajas M.B., Zhang Y., Kumar S., Sharma P.K., Gallos G, & Emala C.W. (2015). Antagonists of the TMEM16A Calcium-Activated Chloride Channel Modulate Airway Smooth Muscle Tone and Intracellular Calcium. Anesthesiology, 123(3), 569-581.
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8

Propranolol Uptake Inhibition Assay

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Cells were pre-incubated with inhibitor at a concentration 100 times higher (400 nM) than that of propranolol to competitively inhibit the membrane channel. Amantadine (Sigma, CAS no. 768-94-5), cimetidine (ICN, CAS no. 51481-61-9), cyclosporine A (Fluka, CAS no. 59865-13-3), MK571 (Tocris Biosciences, CAS no. 115104-28-4), quinidine (Sigma, CAS no. 56-54-2) or verapamil hydrochloride (Fluka, CAS no. 152-11-4) were dissolved in DMSO (0.1% in final solution) and added to the apical (in 0.75 mL freshwater) or basal (in 1.0 mL L-15 medium). Controls and compartments without inhibitor contained 0.1% DMSO. After 1 h, volumes were replaced with 1.5 mL freshwater containing 1 μg L−1 (4 nM) propranolol apically and 2.0 mL L-15 basally, whilst keeping the final concentration of inhibitor at 400 nM throughout. The same sampling procedure as for BTA at time 0, 6 (not for cyclosporine A), 24, 36 and 48 h proceeded and the uptake Papp of propranolol was calculated using Eq. (2). The Papp in all inhibitor-free controls were expressed as a percentage of the mean (100%) and the change in uptake Papp (inhibition) in the presence of inhibitors was expressed as a percentage of this mean control.
Stott L.C., Schnell S., Hogstrand C., Owen S.F, & Bury N.R. (2015). A primary fish gill cell culture model to assess pharmaceutical uptake and efflux: Evidence for passive and facilitated transport. Aquatic Toxicology (Amsterdam, Netherlands), 159, 127-137.
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