ECM910
MDR1 Efflux Assay
The Multidrug Resistance Direct Dye Efflux Assay Kit includes two of the best characterized & most commonly used multidrug resistance ABC transporter substrates, DiOC2(3) & rhodamine 123.
Anmeldenzur Ansicht organisationsspezifischer und vertraglich vereinbarter Preise
Alle Fotos(2)
About This Item
UNSPSC-Code:
12161503
eCl@ss:
32161000
NACRES:
NA.84
Empfohlene Produkte
Qualitätsniveau
Speziesreaktivität (Voraussage durch Homologie)
mammals
Hersteller/Markenname
Chemicon®
Methode(n)
cell based assay: suitable
NCBI-Hinterlegungsnummer
UniProt-Hinterlegungsnummer
Nachweisverfahren
fluorometric
Anwendung
Research Category
Zellstruktur
Zellstruktur
The Multidrug Resistance Direct Dye Efflux Activity Kit is designed to directly measure the functional activity of MDR1, MRP1 and BCRP by assaying for the ability of the cell to extrude fluorescent transport substrates of these proteins. The kit can be used to measure relative amounts of MDR1, MRP and BCRP activity between different cell populations. In addition, dye efflux assays can be used to assess the ability of unlabeled small molecules to serve as transport substrates for MDR1, MRP1 and BCRP and the ability of candidate inhibitors to block the function of these proteins.
Two conveniently formulated solutions of MDR1, MRP1 and BCRP substrate dyes, DiOC2(3) Solution and Rhodamine 123 Solution, are included in the kit to give the investigator flexibility in the substrate specificity of the efflux probe. In addition, a solution of a nonfluorescent substrate of MDR1 and MRP1, Vinblastine Solution, is provided to serve as a competitive inhibitor of dye efflux. DMSO is included as a diluent control for the Vinblastine Solution. Sterile, concentrated RPMI-1640 and BSA are provided as components for buffer for the loading, efflux and washing steps of the procedure. A sterile, concentrated solution of gentamicin, which has been shown to not be a substrate for MDR1 (Mechetner and Roninson, 1992), is supplied as an optional antibiotic for the buffer. Propidium iodide solution is included to identify dead cells and exclude them from the analysis.
The extent of efflux of fluorescent dyes is best analyzed by flow cytometry. The assay can also be incorporated with immunostaining experiments for multicolor analysis of cell surface marker expression and MDR1, MRP1 and BCRP function. Efflux mediated by MDR1, MRP1 and BCRP can also be quantified by analysis under a fluorescence microscope or on a fluorometric plate reader.
The efflux activity of MDR1 and its relatives is highly temperature sensitive. MDR1 functions optimally near 37ºC, but is effectively inactive at 4ºC. MDR1-expressing cells preloaded with MDR1 fluorescent substrates retain the dye and consequently have high fluorescence when incubated at 4ºC (Figure 1, 4ºC, top panels). Conversely, cells incubated at 37ºC more readily efflux the dye and show reduced fluorescence (Figure 1, 37ºC + DMSO, middle panels). The kit also includes a specific inhibitor vinblastine, which is also a substrate for MDR1 and competitively blocks efflux of DiOC2(3) and Rhodamine 123. Inclusion of excess vinblastine in the efflux reaction at 37ºC therefore results in high fluorescence (Figure 1, 37ºC + vinblastine, bottom panels).
For Research Use Only; Not for use in diagnostic or in vivo procedures
Two conveniently formulated solutions of MDR1, MRP1 and BCRP substrate dyes, DiOC2(3) Solution and Rhodamine 123 Solution, are included in the kit to give the investigator flexibility in the substrate specificity of the efflux probe. In addition, a solution of a nonfluorescent substrate of MDR1 and MRP1, Vinblastine Solution, is provided to serve as a competitive inhibitor of dye efflux. DMSO is included as a diluent control for the Vinblastine Solution. Sterile, concentrated RPMI-1640 and BSA are provided as components for buffer for the loading, efflux and washing steps of the procedure. A sterile, concentrated solution of gentamicin, which has been shown to not be a substrate for MDR1 (Mechetner and Roninson, 1992), is supplied as an optional antibiotic for the buffer. Propidium iodide solution is included to identify dead cells and exclude them from the analysis.
The extent of efflux of fluorescent dyes is best analyzed by flow cytometry. The assay can also be incorporated with immunostaining experiments for multicolor analysis of cell surface marker expression and MDR1, MRP1 and BCRP function. Efflux mediated by MDR1, MRP1 and BCRP can also be quantified by analysis under a fluorescence microscope or on a fluorometric plate reader.
The efflux activity of MDR1 and its relatives is highly temperature sensitive. MDR1 functions optimally near 37ºC, but is effectively inactive at 4ºC. MDR1-expressing cells preloaded with MDR1 fluorescent substrates retain the dye and consequently have high fluorescence when incubated at 4ºC (Figure 1, 4ºC, top panels). Conversely, cells incubated at 37ºC more readily efflux the dye and show reduced fluorescence (Figure 1, 37ºC + DMSO, middle panels). The kit also includes a specific inhibitor vinblastine, which is also a substrate for MDR1 and competitively blocks efflux of DiOC2(3) and Rhodamine 123. Inclusion of excess vinblastine in the efflux reaction at 37ºC therefore results in high fluorescence (Figure 1, 37ºC + vinblastine, bottom panels).
For Research Use Only; Not for use in diagnostic or in vivo procedures
The Multidrug Resistance Direct Dye Efflux Assay Kit includes two of the best characterized & most commonly used multidrug resistance ABC transporter substrates, DiOC2(3) & rhodamine 123.
Komponenten
Sterile 5x RPMI-1640 (Part No. 90302) - One bottle containing 200 ml of sterile 5x concentrated RPMI-1640.
Sterile 30% BSA (Part No. 90303) - One bottle containing 35 ml sterile-filtered 30% bovine serum albumin (BSA) in PBS.
Gentamicin Solution, 1000x (Part No. 90353) - One vial containing 1 ml sterile-filtered 50 mg/ml gentamicin.
DiOC2(3) Solution (Part No. 90299) - One vial containing 50 μl of DiOC2(3) (3,3′-diethyloxacarbocyanine iodide) at 1 mg/ml in DMSO.
Rhodamine 123 Solution (Part No. 90300) - One vial containing 0.5 ml of rhodamine 123 at 1 mg/ml in DMSO.
Vinblastine Solution (Part No. 90301) - One vial containing 150 μl of 22 mM vinblastine in DMSO.
DMSO (Part No. 90294) - One vial containing 150 μl DMSO.
Propidium Iodide Stock Solution (Part No. 90296) - One vial containing 1 ml of 50 μg/ml propidium iodide in PBS.
Sterile 30% BSA (Part No. 90303) - One bottle containing 35 ml sterile-filtered 30% bovine serum albumin (BSA) in PBS.
Gentamicin Solution, 1000x (Part No. 90353) - One vial containing 1 ml sterile-filtered 50 mg/ml gentamicin.
DiOC2(3) Solution (Part No. 90299) - One vial containing 50 μl of DiOC2(3) (3,3′-diethyloxacarbocyanine iodide) at 1 mg/ml in DMSO.
Rhodamine 123 Solution (Part No. 90300) - One vial containing 0.5 ml of rhodamine 123 at 1 mg/ml in DMSO.
Vinblastine Solution (Part No. 90301) - One vial containing 150 μl of 22 mM vinblastine in DMSO.
DMSO (Part No. 90294) - One vial containing 150 μl DMSO.
Propidium Iodide Stock Solution (Part No. 90296) - One vial containing 1 ml of 50 μg/ml propidium iodide in PBS.
Lagerung und Haltbarkeit
Kit components should be stored at 2-8°C upon arrival. Product is stable for 1 year after receipt. DiOC2(3) Solution, Rhodamine 123 Solution, Vinblastine Solution, and DMSO will freeze at 2-8°C, and can be thawed and refrozen at least 5 times. Protect DiOC2(3), Rhodamine 123 and Propidium Iodide Solutions from light.
Precautions
Propidium Iodide, Gentamicin, Vinblastine, Rhodamine 123 and DiOC2(3) may have toxic effects if handled improperly. DMSO is absorbed through the skin and can facilitate internalization of toxic agents. Use caution when handling each of these components. Standard precautions include wearing gloves and a lab coat.
Precautions
Propidium Iodide, Gentamicin, Vinblastine, Rhodamine 123 and DiOC2(3) may have toxic effects if handled improperly. DMSO is absorbed through the skin and can facilitate internalization of toxic agents. Use caution when handling each of these components. Standard precautions include wearing gloves and a lab coat.
Rechtliche Hinweise
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Haftungsausschluss
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Signalwort
Danger
H-Sätze
Gefahreneinstufungen
Muta. 2 - Resp. Sens. 1 - Skin Sens. 1
Lagerklassenschlüssel
10 - Combustible liquids
Flammpunkt (°F)
188.6 °F
Flammpunkt (°C)
87 °C
Analysenzertifikate (COA)
Suchen Sie nach Analysenzertifikate (COA), indem Sie die Lot-/Chargennummer des Produkts eingeben. Lot- und Chargennummern sind auf dem Produktetikett hinter den Wörtern ‘Lot’ oder ‘Batch’ (Lot oder Charge) zu finden.
Besitzen Sie dieses Produkt bereits?
In der Dokumentenbibliothek finden Sie die Dokumentation zu den Produkten, die Sie kürzlich erworben haben.
Parveen Abidi et al.
Journal of lipid research, 47(10), 2134-2147 (2006-08-04)
Our previous studies have identified berberine (BBR), an alkaloid isolated from the Chinese herb huanglian, as a unique cholesterol-lowering drug that upregulates hepatic low density lipoprotein receptor (LDLR) expression through a mechanism of mRNA stabilization. Here, we demonstrate that the
Yanrong Qian et al.
Cancers, 12(12) (2020-12-10)
Growth hormone (GH) and the GH receptor (GHR) are expressed in a wide range of malignant tumors including melanoma. However, the effect of GH/insulin-like growth factor (IGF) on melanoma in vivo has not yet been elucidated. Here we assessed the
Archana Thakur et al.
Oncoimmunology, 10(1), 1930883-1930883 (2021-06-15)
In this study, we investigated the ability of bispecific antibody armed activated T cells to target drug resistant pancreatic cancer cells and whether or not "priming" these resistant cancer cells with bispecific antibody armed activated T cells could enhance subsequent
Magdalena Kasendra et al.
eLife, 9 (2020-01-15)
Induction of intestinal drug metabolizing enzymes can complicate the development of new drugs, owing to the potential to cause drug-drug interactions (DDIs) leading to changes in pharmacokinetics, safety and efficacy. The development of a human-relevant model of the adult intestine
Gad D Vatine et al.
Cell stem cell, 24(6), 995-1005 (2019-06-08)
The blood-brain barrier (BBB) tightly regulates the entry of solutes from blood into the brain and is disrupted in several neurological diseases. Using Organ-Chip technology, we created an entirely human BBB-Chip with induced pluripotent stem cell (iPSC)-derived brain microvascular endothelial-like cells
Unser Team von Wissenschaftlern verfügt über Erfahrung in allen Forschungsbereichen einschließlich Life Science, Materialwissenschaften, chemischer Synthese, Chromatographie, Analytik und vielen mehr..
Setzen Sie sich mit dem technischen Dienst in Verbindung.