The number of apoptotic cells following treatment with plumbagin entrapped in transferrin‐bearing liposomes was determined using BD Pharmingen® FITC Annexin V apoptosis detection kit (BD Biosciences), as described in the manufacturer's instructions. Cells were seeded in 6‐well plates at a density of 2 × 105 cells per well and grown for 24 h before being treated with plumbagin (1 μg per well) entrapped in Tf‐bearing liposomes, control liposomes, or in solution. After 4 h treatment, the cells were harvested and centrifuged at 2000 rpm (370 g) for 5 min using an IEC Micromax® centrifuge (ThermoFisher Scientific). Subsequently, the cell pellets were resuspended in 200 μL 1× Annexin V Binding Buffer (10× of the buffer containing 0.1‐M Hepes/NaOH (pH 7.4), 1.4‐M NaCl, and 25‐mM CaCl2). Cell suspension (100 μL) was then transferred to a 5‐mL culture tube, followed by 5 μL of Annexin V‐FITC labelling reagent and 5 μL of propidium iodide and incubated for 15 min at 20°C protected from light. After incubation, 400 μL of Annexin V Binding Buffer was added to each tube before analysis of apoptosis using a FACSCanto® flow cytometer (BD Biosciences). Ten thousand cells (gated events) were counted for each sample. The results were reported as percentages of specific cell populations (live cells, cells in early apoptosis, late apoptosis, and necrosis).
Iec micromax
Manufactured by Thermo Fisher Scientific
Sourced in United States
The IEC Micromax® is a compact, high-speed centrifuge designed for a variety of laboratory applications. It features a brushless motor and electronic controls for precise speed and time settings. The centrifuge accommodates a range of sample volumes and rotor configurations to meet the needs of different laboratory workflows.
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Lab products found in correlation
2 protocols using iec micromax
1
Plumbagin-Induced Apoptosis Quantification
2
Pharmacokinetics of RGD-ACM Liposomes
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The rats were randomized into two groups (n=6 each) to receive either intravenous RGD-ACM liposomes at a dose of 8 mg/kg or the same intravenous dose of ACM liposomes as a control. For the pharmacokinetic studies, 0.5 mL samples of whole blood were collected from both groups at 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 12, and 24 hours post-dosing and immediately centrifuged at 4,000 rpm for 10 minutes to obtain plasma samples. The supernatant plasma (0.25 mL) was transferred to new tubes and stored at −80°C until analysis. Pharmacokinetic parameters were calculated using Microsoft Office Excel 2013.
Next, 100 μL of the plasma sample was transferred to a 10 mL plastic test tube together with 10 μL of internal standard solution (quercetin, 1 mg/mL). After shaking for 30 seconds using a 5432 vortex mixer (Eppendorf, Hamburg, Germany), 3 mL of ethyl acetate was added and the mixture was vortexed for a further 2 minutes. After centrif ugation at 3,000 g for 10 minutes (IEC Micromax, Thermo Scientific, Rockford, IL, USA), the upper organic layer was quantitatively transferred to a 10 mL glass tube and evaporated to dryness using an evaporator at 40°C. The residue was reconstituted in 100 μL of methanol, and then vortex-mixed. After centrifugation at 2,000 g for 5 minutes, a 20 μL aliquot of the solution was injected into the high-performance liquid chromatography system for analysis.
Next, 100 μL of the plasma sample was transferred to a 10 mL plastic test tube together with 10 μL of internal standard solution (quercetin, 1 mg/mL). After shaking for 30 seconds using a 5432 vortex mixer (Eppendorf, Hamburg, Germany), 3 mL of ethyl acetate was added and the mixture was vortexed for a further 2 minutes. After centrif ugation at 3,000 g for 10 minutes (IEC Micromax, Thermo Scientific, Rockford, IL, USA), the upper organic layer was quantitatively transferred to a 10 mL glass tube and evaporated to dryness using an evaporator at 40°C. The residue was reconstituted in 100 μL of methanol, and then vortex-mixed. After centrifugation at 2,000 g for 5 minutes, a 20 μL aliquot of the solution was injected into the high-performance liquid chromatography system for analysis.
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