For total deoxyribonucleic acid (DNA) analysis, cells were washed in PBS and treated with 70% methanol to kill them. Further on, Ethidium Homodimer 4 μM ethidium homodimer in PBS (Thermo Fisher, Cat.# E3599) was added. After 30 min of incubation in the dark, the data were collected (Ex 530 mm, em 645 nm) in a Biotek FL-600 plate reader. For mitochondrial metabolism activity measurement, the MTT-based CellTiter 96® Non-Radioactive Cell Proliferation Assay Kit was used (Promega, Cat.# G4000) following the manufacturer’s instructions (readout at 590 nm and 630 nm in a Biotek FL-600 plate reader).
Fl600 plate reader
Manufactured by Agilent Technologies
Sourced in United States
The FL600 plate reader is a fluorescence microplate reader designed for a variety of research and analytical applications. It is capable of performing fluorescence intensity measurements in microplates.
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Lab products found in correlation
6 protocols using fl600 plate reader
1
Quantifying DNA and Mitochondrial Activity
2
Fibrinogen-Coated Assay for Cell Adhesion
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150 ng fibrinogen in bicarbonate buffer (50 mM, pH 9.2) was coated onto each 96-well followed by incubating at 37 °C for 3–4 h. 1% (w/v) BSA was used to block the non-specific sites at 37 °C for 0.5 h. Coated plate was washed twice before adding cells. Transfected 293 T were labeled by 1 µg/ml BCECF (Invitrogen) at 37 °C for 20 min, followed by transferring to coated plate and incubating at 37 °C for 30 min with or without activator (0.5 mM Mn2+) or blocker (mAb 7E3). The fluorescence signal which indicates the ligand-bound cells of each well was measured using a FL600 plate reader (Bio-Tek instruments, Winooski, USA).
3
Evaluating Alpinetin's Modulation of CYP1A1
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EL-4 cells were seeded into 6-well plates at a density of 1 × 106 cells/mL, and treated with alpinetin (3, 10, 30 μM), siAhR, CH223191 (10 μM), siAhR + alpinetin (30 μM), CH223191 + alpinetin (30 μM) and TCDD (5 nM) for 24 h, and supernatant was collected. Then, activity of CYP1A1 was detected by using an EROD enzyme kit, fluorescence intensity was measured by using a FL600 plate reader with excitation at 530 nm and emission at 590 nm (Biotek, Winooski, VT, USA).
4
Quantifying Oxidative Stress in Cells
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Cells were seeded at 5,000 cells per well in a 96-well plate 15 to 18 hours before the start of the treatment. Aldi-6 or Alda-89 was prepared in DMSO/PBS mix and cisplatin was prepared fresh by adding 1 mg of cisplatin into 1 ml of saline. TritonX-100 Cells were treated two times with the compounds on days 1 and 2 and then on day 4, were washed with PBS. Cells were incubated with 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA, Oxi-select kit, STA-342, Cell Biolabs, San Diego, CA) (20 μM-1 mM) at 37°C in the dark 30 min to 1 hr. Cells were then washed again with PBS and lysed. 2′,7′-dichlorodihydrofluorescein (DCF) fluorescence was measured within 30 minutes using a BioTek FL-600 plate reader (BioTek Instruments, Winooski, Vt., USA) at 485 nm excitation and 530 nm emission wavelengths. Data were expressed in nM of DCF as calculated from the standard curves.
5
Sema4D-Induced CYP1A1 Activity
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EL-4 cells were plated into 6-well plates at a density of 1 × 106 cells/ml; stimulated with Sema4D, anti-PlexinB1 antibody, anti-PlexinB2 antibody, anti-CD72, CH223191, and FICZ either alone or in combination; and incubated for 24 h. The supernatant was then collected. Then, CYP1A1 activity was measured with an EROD enzyme assay, as previously described (28 (link)). Fluorescence intensity was detected by using a FL600 plate reader (Biotek, Winooski, VT, United States), with excitation at 530 nm and emission at 590 nm.
6
Sema4D-Induced CYP1A1 Activity
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The EL-4 cells were plated into 6-well plates at a density of 1 × 10 6 cells/mL, and stimulated with Sema4D, anti-PlexinB1, anti-PlexinB2, anti-CD72, CH223191, and FICZ either alone or in combination, and incubated for 24h, and supernatant was collected. Then, CYP1A1 activity was measured by using the EROD enzyme assay, as previously described [25] . Fluorescence intensity was detected by using a FL600 plate reader (Biotek, Winooski, VT, USA), with excitation at 530 nm and emission at 590 nm.
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