rhIDO1 activity was determined as follows. In brief, the standard reaction mixture (200 μL) contained 50 mM potassium phosphate buffer (KPB) (pH 6.5), 20 mM ascorbic acid (neutralized with NaOH and HCl) (Sigma Aldrich), 100 μg/mL catalase (Sigma Aldrich), 10 μM methylene blue (Alfa Aesar, Heysham, Lancashire, UK), 100 µM l -tryptophan (Sigma Aldrich), 50 nM rhIDO1 (Origene, Bologna, Italy), and dimethyl sulfoxide (DMSO) solution of the compound (4 μL). The reaction was carried out at 37 °C for 60 min and stopped by the addition of 40 μL of 30% (w/v) CCl3COOH. After heating at 50 °C for 15 min, the reaction mixture was centrifuged at 1500× g for 10 min. The supernatant (150 μL) was transferred into a well of a 96-well microplate and mixed with 150 μL of 2% (w/v) p-dimethylaminobenzaldehyde (Ehrlich’s reagent) in acetic acid. The yellow pigment derived from kynurenine was measured at 490 nm using an Ultramark Microplate Imaging System (Bio-Rad, Hercules, CA, USA). IC50 values were calculated from concentration-response curves obtained in at least three different experiments run in triplicate.
Ultramark microplate imaging system
Manufactured by Bio-Rad
Sourced in United States
The Ultramark Microplate Imaging System is a compact and versatile device designed for imaging and analyzing microplates. It utilizes a high-resolution camera and advanced optics to capture images of samples in microplate formats. The system is capable of performing a range of imaging techniques, including absorbance, fluorescence, and luminescence measurements, making it suitable for various applications in life science research and diagnostic laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Celltiter 96 aqueous one solution reagent, by Promega (2 mentions)
Methylene blue, by Thermo Fisher Scientific (1 mentions)
Catalase, by Merck Group (1 mentions)
L tryptophan, by Merck Group (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Facsaria cell sorter, by BD (1 mentions)
Ddh2o, by Merck Group (1 mentions)
7 protocols using ultramark microplate imaging system
1
Determination of rhIDO1 Activity
2
Cell Viability and Cell Cycle Analysis of MMC-Treated U2OS Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
U2OS cell line was transfected with the siControl, siDNAH2, or siFANCA, respectively, and then treated by the different concentrations of MMC (0, 50, 100, 150, 200, 250 ng/mL in ddH2O, Sigma) for 24 h. After the adding of CellTiter 96 Aqueous One Solution Reagent (Promega) for 2 h, cellular viability was measured using an UltraMark Microplate Imaging System (Bio Rad, Hercules, CA) following the manufacturer's protocol. In addition, under the treatment of 100 ng/mL MMC or not, a flow cytometric analysis was performed to analyze the cell cycle status, according to the manufacturer's protocol (BD FACS Aria TM cell sorter). The percentage of the G2/M phase was calculated, respectively.
3
Quantification of Cytokine Levels by ELISA
4
Cell Proliferation Assay with siRNAs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells transfected with control, AURKA and AURKB siRNAs were treated with CellTiter 96® Aqueous One Solution Reagent (Promega, Madison, WI) in accordance with the manufacturer's protocol. After 1 h, proliferation of the cells was measured by absorbance at 490 nm using an UltraMark Microplate Imaging System (Bio Rad, Hercules, CA). Results were presented as the mean ± standard deviation of triplicate determinations.
5
Inhibition of IDO1 Enzymatic Activity by VS9
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The effects of VS9 on the enzymatic activity of IDO1 were determined using the IDO1 inhibitor screening kit (BioVision Incorporate Milpitas CA, USA), according to manufacturer instructions. VS9 (10 μM) was added to complete assay reaction buffer and incubated for 45 min at 37 °C. The reaction was stopped by the addition of 30 % (w/v) CCl3COOH. After heating at 50 °C for 15 min, the reaction mixture was centrifuged at 1500 g for 10 min. The supernatant was transferred into a well of a 96‐well microplate and mixed (1 : 1 ratio) with of 2 % (w/v) p‐dimethylaminobenzaldehyde (Ehrlich's reagent) in acetic acid. The yellow pigment derived from kynurenine was measured at 490 nm using an Ultramark Microplate Imaging System (Bio‐Rad). A positive inhibition control, included in the kit, was added. The results are expressed as mean±SEM of three different experiments run in triplicate.
6
Quantifying Betacyanin Content in Amaranth
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Leaf, stem and root tissue samples of the three genotypes of A. hypochondriacus plants subjected to the different stress treatments described above, together with those obtained from the respective controls, were homogenized in liquid nitrogen. Betacyanins were extracted in water and the pigment content in the solutions was determined by spectrophotometrical determination at 536 nm, using an Ultramark Microplate Imaging System (Bio-Rad Laboratories, Hercules CA, USA). The betacyanin content of the plant aqueous extracts was estimated using the molar extinction coefficient for amaranthine (5.66×104 L mol−1 cm−1;[22] ) and a MW of 726.6.
7
Antimicrobial Efficacy of CoLig Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The susceptibility of S. aureus and P. aeruginosa to CoLig NPs was determined in 96-well microtiter plates through the broth dilution method as previously described [47 (link)]. Different dilutions of CoLig NPs were incubated at 37 °C with bacterial suspensions (5 × 105 CFU/mL) in Nutrient broth. After 24 h, the optical density at 600 nm was measured (Ultramark Microplate Imaging System, Bio-Rad Laboratories S.r.l., Segrate, Italy) to obtain the minimum inhibitory concentration (MIC). As a control, Lig NPs were used.
The morphology of S. aureus and P. aeruginosa treated with CoLig NPs was examined using SEM (Merlin Zeiss, operating at 1 kV). Bacterial cultures were grown in NB overnight and then diluted to an OD600 = 0.01. The suspension was mixed with CoLig NPs to achieve a final concentration of 2.4 mg/mL and transferred to a 48-well plate containing silicon wafers. After 24 h at 37 °C, the liquid was drained, and the bacteria remaining on the wafers were fixed overnight in a 2% paraformaldehyde solution. Finally, the bacteria were dehydrated by incubating the wafers with increasing concentrations of ethanol for 1 h each (25, 50, 75, and 100%). The same process was repeated for untreated bacteria.
The morphology of S. aureus and P. aeruginosa treated with CoLig NPs was examined using SEM (Merlin Zeiss, operating at 1 kV). Bacterial cultures were grown in NB overnight and then diluted to an OD600 = 0.01. The suspension was mixed with CoLig NPs to achieve a final concentration of 2.4 mg/mL and transferred to a 48-well plate containing silicon wafers. After 24 h at 37 °C, the liquid was drained, and the bacteria remaining on the wafers were fixed overnight in a 2% paraformaldehyde solution. Finally, the bacteria were dehydrated by incubating the wafers with increasing concentrations of ethanol for 1 h each (25, 50, 75, and 100%). The same process was repeated for untreated bacteria.
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!