The cells were seeded at a density of 5 × 104 cells/well in 96-well plates and allowed to incubate overnight. Once the cells adhered to the wells, lipopolysaccharide (LPS, 1 μg/mL) was added, both with and without the addition of the isolated compounds. After the treatment of the tested compounds, the Griess reagent method was employed to access the nitrate levels, which consisted of a 1:1 ratio of solution A (1% sulfanilamide) and solution B (5% phosphoric acid with 0.1% naphthyl ethylenediamine-HCl). A microplate reader (VersaMax, Molecular Devices, LLC., San Jose, CA, USA) was used to measure the absorbance at 540 nm. A standard nitrite was used to evaluate the nitrite levels in the samples. Cell viability was assessed using a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. In short, 3T3-L1 adipocytes were seeded and cultured in DMEM with 10% FBS on 96-well plates. After 24 h of incubation, the cells were treated with the test compounds, which were dissolved in a serum-free medium for one day. Subsequently, 20 µL of the 2 mg/mL MTT solution (Sigma) was added to each well and left to incubate in the dark for 4 h. After discarding the supernatant, formazan was dissolved in DMSO, and the absorbance was measured at 550 nm using a microplate reader (VersaMax, Molecular Devices, LLC., San Jose, CA, USA).
Versamax
Manufactured by Molecular Devices
Sourced in United States, United Kingdom, Japan, China, Germany, Brazil
The VersaMax is a microplate reader designed for a variety of assays. It can measure absorbance, fluorescence, and luminescence in 96- and 384-well microplates. The device provides consistent and reliable performance for researchers in a variety of applications.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (22 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (13 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (12 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (10 mentions)
Softmax pro, by Molecular Devices (9 mentions)
Softmax pro software, by Molecular Devices (8 mentions)
Oil red o, by Merck Group (6 mentions)
Mtt solution, by Merck Group (6 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (6 mentions)
Elisa kit, by R&D Systems (6 mentions)
Bovine serum albumin (bsa), by Merck Group (6 mentions)
Acetylated prothrombin, by Enzyme Research (6 mentions)
Factor xa, by Enzyme Research (6 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (5 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (5 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (5 mentions)
Ez cytox cell viability assay kit, by Daeil Lab Service (4 mentions)
Anti mouse igg hrp, by Southern Biotech (4 mentions)
Elisa kit, by BioLegend (4 mentions)
Griess reagent, by Merck Group (4 mentions)
1 039 protocols using versamax
1
Evaluating Anti-Inflammatory Effects of Compounds
2
Cytotoxicity of DOX-loaded Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The A549 tumor cells (5 × 103 cells/well) were seeded onto 96-well plates and stabilized for 12 h. The A549 cells were then incubated with various concentrations (0, 0.1, 1, 10, 100 and 500 μg/mL of DOX) of free DOX, CNPs and DOX-CNPs for 24 h. To measure cell viability, 10% (v/v) CCK-8 solution was added to each well, followed by further incubation for 1 h at 37 °C. The absorbance at 450 nm was measured using a microplate reader (VERSAmax™, Molecular Devices Corp., Sunnyvale, CA, USA).
To analyze the cell viability after HIFU treatment, A549 cells were incubated with CNPs and DOX-CNPs (100 μg/mL of DOX) for 24 h. These A549 cells were exposed to HFU in destruction mode (power: 10 MHz, mechanical index: 0.235) for 5 min, followed by further incubation for 24 h at 37 °C in a 5% CO2 incubator. Finally, the A549 cells were washed twice with DPBS. Subsequently, 10% (v/v) CCK-8 solution was added to each well, followed by further incubating for 1 h at 37 °C. The absorbance at 450 nm was measured using a microplate reader (VERSAmax™, Molecular Devices Corp., Sunnyvale, CA, USA).
To analyze the cell viability after HIFU treatment, A549 cells were incubated with CNPs and DOX-CNPs (100 μg/mL of DOX) for 24 h. These A549 cells were exposed to HFU in destruction mode (power: 10 MHz, mechanical index: 0.235) for 5 min, followed by further incubation for 24 h at 37 °C in a 5% CO2 incubator. Finally, the A549 cells were washed twice with DPBS. Subsequently, 10% (v/v) CCK-8 solution was added to each well, followed by further incubating for 1 h at 37 °C. The absorbance at 450 nm was measured using a microplate reader (VERSAmax™, Molecular Devices Corp., Sunnyvale, CA, USA).
3
Serum ALT and Creatinine Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum ALT level was measured using an alanine aminotransferase (ALT or SGPT) activity colorimetric/fluorometric assay kit (#K752-100, BioVison, CA, USA). The optical density at 570 nm was measured using a microplate reader (VersaMax with SoftMax Pro software, Molecular Devices, CA, USA).
Serum creatinine levels were measured using a creatinine colorimetric/fluorometric assay kit (#K625-100, BioVison, CA, USA). The optical density at 570 nm was measured using a microplate reader (VersaMax with SoftMax Pro software, Molecular Devices, CA, USA).
Serum creatinine levels were measured using a creatinine colorimetric/fluorometric assay kit (#K625-100, BioVison, CA, USA). The optical density at 570 nm was measured using a microplate reader (VersaMax with SoftMax Pro software, Molecular Devices, CA, USA).
4
Homocysteine-Induced Endothelial Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Viability of HRECs after treatment with Hcy (100, 50 and 20 µM) was determined using MTT Cell Proliferation Assay Kit (Invitrogen, Grand Island, NY, USA). HRECs were grown on 96-well plates at a density of 2 × 104 cells/well in phenol red free medium and incubated with different concentrations of Hcy for 24 hours. The medium was removed and the cells were treated with 20 μL of MTT (5 mg/mL) and incubated for 4 h hours at 37 °C prior to the addition of 100 μL of DMSO. The resulting formazan product was measured spectrophotometrically at 540 nm using a microplate reader (VERSA max, Molecular Devices, Sunnyvale, CA, USA).
Caspase-3/7 activity in homocysteine treated HRE cells was measured using the Apo-ONE Homogenous caspase-3/7 system according to the manufacturer’s instructions (Promega). The intensity of the emitted fluorescence was determined at an excitation wavelength range of 485 ± 20 nm and an emission wavelength range of 530 ± 25 nm using microplate reader (VERSA max, Molecular Devices, Sunnyvale, CA, USA). Caspase-3/7 activity was expressed as net fluorescence intensity (RFU). Data shown are means ± SD of three different independent experiments.
Caspase-3/7 activity in homocysteine treated HRE cells was measured using the Apo-ONE Homogenous caspase-3/7 system according to the manufacturer’s instructions (Promega). The intensity of the emitted fluorescence was determined at an excitation wavelength range of 485 ± 20 nm and an emission wavelength range of 530 ± 25 nm using microplate reader (VERSA max, Molecular Devices, Sunnyvale, CA, USA). Caspase-3/7 activity was expressed as net fluorescence intensity (RFU). Data shown are means ± SD of three different independent experiments.
5
Evaluating EGFR Inhibitor Potency
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BaF3‐EGFR19del/T790M/C797S, BaF3‐EGFRL858R/T790M/C797S, or BaF3‐EGFR19del/T790M cells were plated in 96‐well plates at a density of 5000 cells per well and cultured overnight. Different concentrations of compounds were added, and the cells were incubated at 37°C in a CO2 incubator for 72 hours. Then, the cell growth was measured using Cell Counting Kit‐8 (CCK8; #AC11L057, Life iLab) and multiwell spectrophotometer (VERSA max™, Molecular Devices) at an absorbance of 450 nm.
PC‐9 cells or PC‐9‐OR cells were plated in a 96‐well culture plate at a density of 2000 cells per well and cultured overnight. The compounds in different concentrations were added, and then the cells were cultured for 72 hours. Then, sulforhodamine B (SRB, #S9012, Sigma‐Aldrich) assay was performed according to standard protocols, and the results were acquired using a multiwell spectrophotometer (VERSA max™, Molecular Devices) at an absorbance of 560 nm.
PC‐9 cells or PC‐9‐OR cells were plated in a 96‐well culture plate at a density of 2000 cells per well and cultured overnight. The compounds in different concentrations were added, and then the cells were cultured for 72 hours. Then, sulforhodamine B (SRB, #S9012, Sigma‐Aldrich) assay was performed according to standard protocols, and the results were acquired using a multiwell spectrophotometer (VERSA max™, Molecular Devices) at an absorbance of 560 nm.
6
Comparative Evaluation of MSC Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Vybrant® MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] Cell Proliferation Assay was used for the comparative evaluation of proliferative potential of MSCs. Briefly, 1 × 103 MSCs were cultured in 96 well plate and every other day cells were incubated with 20 μl MTT solution (5 mg/ml) for 4 hr. After incubation, the supernatant was removed and 50 μl dimethyl sulfoxide (DMSO) was added. The product formed was then transferred to 96 well plate and the absorbance was measured at 540 nm using microplate reader (VersaMax™, Molecular Devices, CA, USA). The senescence-associated β-galactosidase activity (SA-β-gal) was measured using β-galactosidase Assay Kit following manufacturer's protocol. Briefly, cells at 70-80% confluence were harvested by treating with TrypLE and washed twice with DPBS, and a total of 5 × 105 cells were treated with 200 μl M-PER reagent and then gently agitated for 10 min. After being centrifuged at 27,000 ×g for 15 min to remove the debris of cell lysis, 50 μl of the supernatant was transferred into a 96-well microplate. The cell extract was incubated at 37°C for 30 min with 50 μl of β-galactosidase reagent and the absorbance was measured at 405 nm using microplate reader (VersaMax™, Molecular Devices, CA, USA).
7
Evaluating GBE's Effects on HT-22 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HT-22 cells, a murine normal hippocampal neuronal cell line, were obtained from Millipore and maintained at 37°C with 5% CO2. After HT-22 cells (1×104 cells/well) were grown for 24 h, the cells were treated with GBE (0.01–100 µg/ml) for 72 h to examine the effect of GBE on the viability of HT-22 cells. In another experiments, GBE-pretreated HT-22 cells (0.01–100 µg/ml, 0.5 h) were subsequently exposed to 5 mM of glutamate or 500 µM of hydrogen peroxide (H2O2) for 12 h. Cell viability was measured at wavelength of 450 nm using EZ-Cytox cell viability assay kit (Daeil Labservice) and automated microplate reader (VersaMax™; Molecular Devices). The cell viability was calculated as relative to the untreated control cells.
8
Measuring Intracellular cAMP in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
OSNs were plated at a density of 2 × cells/mL in laminin-coated plates, and OR-expressing Hana3A cells were plated at a density of /mL in poly-D-lysine–coated plates 24 h prior to the assay. The intracellular cAMP levels were measured by means of the enzyme immunoassay Parameter® kit (R&D Systems, Minneapolis, MN, USA). The principle of the cAMP assay is competitive binding. The cAMP within the sample competes with a fixed amount of HRP-labeled cAMP for sites on the monoclonal antibody. cAMP was quantified at 450 nm.
In case of HepG2 cells, they were lysed with 0.1 M HCl at 500 μl/well. The lysates were immediately processed with the Direct cAMP ELISA kit (Enzo Life Sciences, Farmingdale, NY, USA) to measure the intracellular concentration of cAMP. The data were analyzed on a microplate reader (Versa Max, Molecular devices, CA, USA) at 405 nm.
In case of HepG2 cells, they were lysed with 0.1 M HCl at 500 μl/well. The lysates were immediately processed with the Direct cAMP ELISA kit (Enzo Life Sciences, Farmingdale, NY, USA) to measure the intracellular concentration of cAMP. The data were analyzed on a microplate reader (Versa Max, Molecular devices, CA, USA) at 405 nm.
9
Evaluating Cisplatin-Induced Cytotoxicity in HK-2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To evaluate the cell viability, HK-2 cells were seeded onto 96-well plates and then treated with cisplatin in serum-free media for 24 h. The cell viabilities were determined using the Ez-cytox assay kit (DOGEN, Seoul, Korea). After removing the culture media, the cells were stained with a WST assay reagent for 1 h at 37 °C in a 5% CO2 incubator. After seeding of HK-2 cells into 6-well plates, the cell were treated with cisplatin, cisplatin + ten herbal medicines for 24 h. Cell culture supernatants were applied into NGAL, KIM-1 pre-coated plates and according to the manufacturer’s instructions (Cloud-Clone Corp, USA, cat. SEB388Hu, SEA785Hu, respectively). To determine whether low and high doses of cisplatin induce apoptosis or necrosis, cells treated with 400 μM of cisplatin were assessed at 2, 4, and 6 h, and cells treated with 10 μM of cisplatin were assessed at 6, 24, and 48 h. The absorbance was determined in lysates at 450 nm using an ELISA reader (VERSA Max; Molecular Devices, Sunnyvale, CA, USA).
10
Quantifying Adipocyte Lipid Content
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fully differentiated 3T3-L1 cells were washed twice with PBS and fixed in 4% paraformaldehyde for 1 h. Cells were stained with 3 g/L of Oil Red O (Sigma Chemical, St. Louis, MO) in 60% isopropanol at room temperature for 10 min and washed extensively with distilled water. Pictures were taken using a microscope (Axiovert 40CFL; Olympus, Germany). In addition, stained Oil Red O dye was extracted with isopropanol and collected, and the absorbance (O.D. 500 nm) was measured by microplate reader (Versamax; Molecular Devices Corporation, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!