Mk571

Manufactured by Selleck Chemicals

Sourced in United States, China

MK571 is a lab equipment product that functions as a multifunctional protein that acts as an inhibitor of the organic anion transport protein MRP1 (ABCC1). It is commonly used in research settings to study cellular transport mechanisms and their modulation.

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Lab products found in correlation

7 protocols using mk571

Labeling and Conjugation of APR-246 Compounds

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APR‐246 (2‐hydroxymethyl‐2‐methoxymethyl‐1‐azabicyclo[2,2,2]octan‐3‐one), ¹⁴C‐labeled APR‐246 and glutathione‐conjugated MQ (GS‐MQ) were obtained from Aprea Therapeutics AB (Stockholm, Sweden). APR‐246 was dissolved in either DMSO or water. MK‐571 (final concentration to cells 5–20 μM) and Sulfasalazine (final concentration to cells 100–200 μM) were from Merck (Germany). MK‐571 for treatment of esophageal cancer cells in vitro and in vivo was from Selleckchem (Houston, TX) or for colo‐PDOs from Cayman Chemical (USA) in 0.9% saline. Reversan (final concentration to cells 10–20 μM) was from Santa Cruz Biotechnology, sc‐296262 (USA). Inhibitors were dissolved in DMSO. The percentage of DMSO during drug exposure never exceeded 0.2% for cell lines and 0.5% for eso‐PDOs. Doxycycline and chemicals described hereafter were all from Merck (Germany).

Ceder S., Eriksson S.E., Cheteh E.H., Dawar S., Corrales Benitez M., Bykov V.J., Fujihara K.M., Grandin M., Li X., Ramm S., Behrenbruch C., Simpson K.J., Hollande F., Abrahmsen L., Clemons N.J, & Wiman K.G. (2020). A thiol‐bound drug reservoir enhances APR‐246‐induced mutant p53 tumor cell death. EMBO Molecular Medicine, 13(2), e10852.

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Cytotoxic Agents and Cell Lines

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The J45.01 T-cell line and Toledo B-cell line were obtained from American Type Culture Collection (Manassas, VA). KBM3/Bu250⁶ is a busulfan-resistant AML cell line established in our laboratory as previously described [10 (link)]. MOLM14 is an AML cell line obtained from Dr. Michael Andreeff's laboratory (UT MD Anderson Cancer Center, Houston, TX). All cells were cultured in RPMI 1640 (Mediatech, Manassas, VA) supplemented with 10% heat-inactivated fetal bovine serum (FBS: Sigma-Aldrich, St. Louis, MO) and 100 U/mL penicillin and 100 μg/mL streptomycin (Mediatech) at 37°C in a humidified atmosphere of 5% CO₂ in air. 5-Carboxyfluorescein diacetate acetoxymethyl ester (5-CFDA) was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA). Verapamil and MK571 were purchased from Selleckchem (Houston, TX). Chlorambucil, busulfan, melphalan, bendamustine, cyclophosphamide, ifosfamide, ketoconazole, posaconazole, fluconazole, itraconazole, metronidazole, ethacrynic acid, everolimus, sirolimus, phenytoin, levetiracetam, and buthionine sulphoximine (BSO) were obtained from Sigma-Aldrich Corp. (St. Louis, MO). 4-Hydroperoxycyclophosphamide (4-HC) and 4-hydroperoxyifosfamide (4-HI) were generous gifts from from Dr. Scott Rowley (Hackensack University Medical Center, Hackensack, New Jersey), and Dr. Robert F. Struck (Southern Research Institute, Birmingham, Alabama) respectively.

Valdez B.C., Hassan M, & Andersson B.S. (2017). Development of an assay for cellular efflux of pharmaceutically active agents and its relevance to understanding drug interactions. Experimental hematology, 52, 65-71.

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Transporter-Mediated Paclitaxel Resistance Study

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Paclitaxel, docetaxel, and doxorubicin were purchased from Sigma-Aldrich (St Louis, MO, USA). MK-571 and puromycin were obtained from Selleck (Shanghai, China). All drugs were dissolved in DMSO. Cell Counting Kit-8 (CCK8) was purchased from Pepro Tech (Wuhan, China). Pig kidney cells (LLC-PK1) were purchased from Wuhan Bafeier Biological Co. Ltd (Wuhan, China). LLC-PK1 stable transfected with empty vector (pcDNA3.1), ABCC2_1249G wild-type allele and ABCC2_1249A variant allele were obtained from Biofavor Biotech (Wuhan, China). Dulbecco’s modified eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from PAN-Biotech (Aidenbach, Germany). MRP2 and β-actin monoclonal antibody were purchased from Abcam (Cambridge, UK). HRP-conjugated IgG was also obtained from Abcam (Cambridge, UK). BCA protein assay kit was obtained from Innova Biosciences (Cambridge, UK).

Lian G., Yuan J, & Gao Y. (2020). In vitro Transport Ability of ABCC2 (G1249A) Polymorphic Variant Towards Anticancer Drugs. OncoTargets and therapy, 13, 1413-1419.

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Detailed Drug Preparation Protocol

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PTX, Dox, Vin, and Ifosfamide (Ifos) were obtained from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Calcein acetoxymethyl ester (Calcein AM) and Green CMFDA were obtained from Abcam (Abcam, Cambridge, MA, USA). Tariquidar, Cyclosporin, and MK-571 were purchased at SelleckChem (Houston, TX, USA). The chemicals were dissolved in dimethyl sulfoxide (DMSO) according to the manufacturer’s recommendations. For in vivo experiments, the stock solution of Dox (1 mg/mL) was prepared in DMSO and diluted before administration in a formulation that contained a final concentration of 5% DMSO, 40% PEG-400, 5% Tween 80 (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany), and 50% dd H₂O.

Boichuk S., Bikinieva F., Valeeva E., Dunaev P., Vasileva M., Kopnin P., Mikheeva E., Ivoilova T., Mustafin I, & Galembikova A. (2023). Establishment and Characterization of Multi-Drug Resistant p53-Negative Osteosarcoma SaOS-2 Subline. Diagnostics, 13(16), 2646.

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Oxidative Stress Modulation in Cell Lines

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p-Arsanilic acid, 70% ammonium thioglycolate solution, and 1,3-propanedithiol, L-buthionine-(S,R)-sulfoximine (BSO) and β-nicotinamide adenine dinucleotide (NADH) were purchased from Aladdin (Shanghai, China). RPMI 1640 Medium, Dulbecco’s modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were obtained from GIBCO (Grand Island, NY, USA). (±)-α-lipoic acid (LA), Hoechst 33342, propidium iodide (PI) and 2′,7′-dichlorfluorescein diacetate (DCFH-DA) were obtained from Sigma-Aldrich (St. Louis, MI, USA). 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was obtained from Amresco (Solon, OH, USA). MK571 was obtained from Selleck (Shanghai, China). Mitochondria Staining Kit (JC-1) and Annexin V-FITC/PI apoptosis kit were purchased from MultiSciences (Hangzhou, China). Tetramethylrhodamine (TMRM) was obtained from HEOWNS (Tianjin, China). Human Cyt C ELISA Kit and Human PDH E2 ELISA Kit were obtained from Elbio (Shanghai, China). RIPA buffer, BCA Protein Assay Kit, ATP Assay Kit, and dihydroethidium (DHE) were purchased from Beyotime (Shanghai, China). Human PDHC Assay Kit was purchased from Letter Sail (Nanjing, China). A 10-mM solution of PDT-PAO-F16 was prepared in DMSO and stored at 4 °C. Other common chemicals were of analytical reagent grade from Shenshi (Wuhan, China) and used without further purification.

Liu Y.J., Fan X.Y., Wang A.D., Xia Y.Z., Fu W.R., Liu J.Y., Jiang F.L, & Liu Y. (2019). LDHA Suppression Altering Metabolism Inhibits Tumor Progress by an Organic Arsenical. International Journal of Molecular Sciences, 20(24), 6239.

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Assessing Cell Death and Mitochondrial Function

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Cell death was measured by flow cytometry after annexin V-FITC/ propidium iodide staining, according to manufacturer’s instructions (eBioscience, Inc., San Diego, CA, USA). Cell lines with a high expression of the multidrug efflux transporter P-glycoprotein (P-gp) according to the cellMiner database [15 (link)] were co-treated with a Pgp inhibitor to avoid the bias of drug efflux during the assessment of toxicity. Reactive Oxygen Species (ROS) quantification was performed with 10 µM H2DCF-DA (Abcam, Cambridge, UK) or 1 µM MitoSOX™ Red (Invitrogen, Carlsbad, CA, USA). In total, 1 µM tariquidar and 50 µM MK571 (Selleckchem) were added to the medium to inhibit fluorescent compound efflux by ABC transporters. The mitochondrial membrane potential was measured after incubation of cells in phenol red-free medium with 5 µM JC1 (Invitrogen) and 1 µM tariquidar. Cytochrome c release was measured by flow cytometry using the Cytochrome c Release Apoptosis Assay Kit (Millipore).

Basseville A., Violet P.C., Safari M., Sourbier C., Linehan W.M., Robey R.W., Levine M., Sackett D.L, & Bates S.E. (2022). A Histone Deacetylase Inhibitor Induces Acetyl-CoA Depletion Leading to Lethal Metabolic Stress in RAS-Pathway Activated Cells. Cancers, 14(11), 2643.

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Comprehensive Cancer Drug Protocol

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Olaparib (AZD2281; #S1060), niraparib (MK-4827; #S2741), rucaparib (#AG-014699) veliparib (ABT-888; #S1004), talazoparib (BMN 673; #S7048), phosphate (#S1098), verapamil (CP-16533-1) HCl (#S4202), SN-38 (NK012; #S4908) and MK-571 (#S8126) were purchased from Selleckchem (Munich, Germany). Cisplatin (1 mg/mL), doxorubicin, paclitaxel (6 mg/mL), and topotecan (1 mg/mL) were purchased from Accord Healthcare GmbH (Munich, Germany). Diphenhydramine-hydrochloride (DIPH; D3630-5G), methyl methanesulfonate (MMS; #129925-5G), and treosulfane (Trecondi; #SML1252) were purchased from Sigma Aldrich. Pegylated liposomal doxorubicin (Caelyx) was purchased from Elblandkliniken Stiftung & Co. KG (Riesa, Germany).

Klotz D.M., Schwarz F.M., Dubrovska A., Schuster K., Theis M., Krüger A., Kutz O., Link T., Wimberger P., Drukewitz S., Buchholz F., Thomale J, & Kuhlmann J.D. (2023). Establishment and Molecular Characterization of an In Vitro Model for PARPi-Resistant Ovarian Cancer. Cancers, 15(15), 3774.

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