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.
Fl bopidy
Manufactured by Merck Group
The FL-BOPIDY is a specialized laboratory instrument designed for fluorescence-based analysis. It is capable of detecting and quantifying fluorescent molecules within samples. The core function of the FL-BOPIDY is to provide accurate and reliable fluorescence measurements, enabling researchers and scientists to conduct a wide range of applications in fields such as biochemistry, molecular biology, and analytical chemistry.
Automatically generated - may contain errors
Lab products found in correlation
Mk571, by Merck Group (3 mentions)
Ripa buffer, by Thermo Fisher Scientific (3 mentions)
Bicinchoninic acid (bca), by Thermo Fisher Scientific (3 mentions)
Rhodamine 123, by Merck Group (3 mentions)
Ko143, by Merck Group (3 mentions)
Cm dcfda, by Merck Group (3 mentions)
Synergymx2, by Agilent Technologies (2 mentions)
Cyclosporine a, by Merck Group (1 mentions)
Synergymx2 elisa plate reader, by Agilent Technologies (1 mentions)
3 protocols using fl bopidy
1
Quantifying Efflux Pump Activity in Cells
2
Efflux Pump Inhibitor Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were incubated in the presence of 10µM Rhodamine 123 (P-gp substrate, Sigma), FL-BOPIDY (BCRP substrate, Sigma) or CM-DCFDA (MRP substrate, Sigma) for 1 hour at 37ºC followed by cell lysis using RIPA buffer (ThermoFisher). For assessing the contribution of e ux pump in the drug uptake, cells were pre-incubated for 1 hour in presence of 5µM cyclosporine A (CsA, P-gp inhibitor, Sigma), 1µM Ko143
(BCRP inhibitor, Sigma) or 10µM MK571 (MRPs inhibitor, Sigma) and maintained during the incubation with drug e ux substrate. Following incubation, cells were brie y washed with ice-cold PBS and lysed with RIPA buffer. Fluorescence in cell lysates was assessed using a SynergyMX 2 ELISA plate reader (Bio-Tek, Winooski, VT, USA). Relative uorescence 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 uorescence 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.
(BCRP inhibitor, Sigma) or 10µM MK571 (MRPs inhibitor, Sigma) and maintained during the incubation with drug e ux substrate. Following incubation, cells were brie y washed with ice-cold PBS and lysed with RIPA buffer. Fluorescence in cell lysates was assessed using a SynergyMX 2 ELISA plate reader (Bio-Tek, Winooski, VT, USA). Relative uorescence 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 uorescence 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.
3
Quantification of Efflux Pump Activities
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were incubated in the presence of 10µM Rhodamine 123 (P-gp substrate, Sigma), FL-BOPIDY (BCRP substrate, Sigma) or CM-DCFDA (MRP substrate, Sigma) for 1 hour at 37ºC followed by cell lysis using RIPA buffer (ThermoFisher). For assessing the contribution of efflux pump in the drug uptake, cells were preincubated for 1 hour in presence of 5µM cyclosporine A (CsA, P-gp inhibitor, Sigma), 1µM Ko143 (BCRP inhibitor, Sigma) or 10µM 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 SynergyMX 2 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.
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