In HUVECs, the toxicity of the berries solutions was assessed by lactate dehydrogenase (LDH) release. The enzyme LDH catalyzes the conversion of lactate to pyruvate via NAD+ reduction to NADH; next the diaphorase reduces tetrazolium salt, oxidizing NADH, to a red formazan product that can be spectrophotometrically determined at 490 nm. LDH release from HUVECs treated for 24 h with berries solutions was monitored by collecting aliquots of the medium under the same experimental conditions performed above for cell viability assays, as previously reported [35 (link)]. The decrease in absorbance between the treatment after 24 h and the control was monitored at 37 °C at 490 nm using an AMR-100 Microplate reader (Allsheng).
Amr 100 microplate reader
Manufactured by Allsheng
Sourced in China
The AMR-100 is a microplate reader designed for absorbance measurements. It can read standard 96-well microplates. The device is capable of performing photometric measurements across a wavelength range.
Automatically generated - may contain errors
6 protocols using amr 100 microplate reader
1
Berries Toxicity Evaluation in HUVECs
2
Cell Proliferation Assay with Medicated Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation and viability were evaluated by CCK-8 assay (Dojindo, Minato, Japan). GSMCs were seeded on 96-well plate (100 μL, 3 × 103 cells/well), allowed to be adherent, and then treated with 300 μL diluted medicated serum, including 10% vehicle-treatment, 10% domperidone, 5%, 10%, 20% AuCi or AuCiBup. The experiment was performed in hexaplicate. After incubation with medicated serum for 12 h, 24 h or 36 h, 10 μL CCK-8 reagent was added to each cell and allowed to react for 1 h. The OD value at 450 nm was measured by an AMR-100 microplate reader (Allsheng, Hangzhou, China).
3
Berberrubine Derivatives Inhibit Yeast α-Glucosidase
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Berberrubine derivatives will be assayed for yeast α-glucosidase inhibitory activity. The protocol described by Ramadhan et al. will be used37 (link). Briefly, yeast α-glucosidase (0.1 U/mL) and substrate (1 mM p-nitrophenyl-α-D-glucopyranoside) were dissolved in 0.1 M phosphate buffer (pH 6.9). A 10 μL test sample was pre-incubated with α-glucosidase (40 μL) at 37 °C for 10 min. A substrate solution (50 μL) was then added to the reaction mixture and incubated at 37 °C for an additional 20 min, and terminated by adding 1 M Na2CO3 solution (100 μL). Enzymatic activity was quantified by measuring the absorbance at 405 nm (ALLSHENG AMR-100 microplate reader). The percentage inhibition of activity was calculated as follows: % Inhibition = [(A0 − A1)/A0] × 100, where: A0 is the absorbance without the sample; A1 is the absorbance with the sample. The IC50 value was deduced from the plot of % inhibition versus the concentration of the test sample. Acarbose was used as standard control and the experiment was performed in triplicate.
4
DPPH Radical Scavenging Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DPPH radical scavenging assay was performed according to the previously described method with minor modification [3 (link),15 (link)]. The samples and a positive control, ascorbic acid, were dissolved in DMSO with final concentrations of 6.25, 12.5, 25, 50, 100, and 200 µg/mL. DPPH was dissolved in anhydrous ethanol (EtOH) with a concentration of 0.04 mg/mL. Tested samples (50 µL) were added to 50 µL of fresh DPPH, then kept in room temperature in the dark for 30 min. The optical density (OD) was measured by an AMR-100 microplate reader (Hangzhou Allsheng Instruments, Hangzhou, China) at 517 nm. The EtOH and DMSO were used as a blank and negative control, respectively. The IC50 values were determined by the software of GraphPad Prism 8 (GraphPad Software Inc., San Diego, CA, USA) [3 (link)].
5
Protective Effects of Compounds on Doxorubicin-Induced Cardiotoxicity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H9c2 rat cardiomyocyte cell lines were obtained from the cell bank of the Chinese Academy of Sciences (Shanghai, China). Cells were maintained with Dulbecco's modified Eagle's medium (DMEM) containing 10% foetal bovine serum (Gibco, New Zealand) supplemented with 1% penicillin-streptomycin. H9c2 cells were cultured in a 5% CO2 incubator at 37 °C. H9c2 rat cardiomyocyte cells, seeded at densities of 4.5 × 104 cells/mL into 96-well plates, were pretreated with different concentrations of six weakly retained components and five strongly retained components for 6 h, followed by 2 μM DOX for 18 h to generate the cell injury model. Cell viability was then tested by CCK-8 assay (Beyotime Biotechnology). 10 µL of CCK-8 solution was added to each well and incubated at 37 °C for 2 h. The resulting colour was determined at 450 nm using an AMR-100 microplate reader (Hangzhou Allsheng Instruments Co., Ltd., China). All data are expressed as the means ± standard deviations (SD). The significant differences were assessed by one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison tests.
6
GFLV/ArMV Detection via DAS-ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) [33] (link) was performed using polyclonal antibodies to GFLV/ArMV (Agritest srl, Valenzano, Italy). The samples were also analyzed for ArMV in order to verify the presence of this virus in Sicilian commercial vineyards, as to date it has not been reported. Fivehundred mg of floematic tissue of each sample was mixed and homogenized with 5 mL extraction buffer (37.2 g TRIS-HCl, 32 g TRIS-base, 8 g NaCl, 20 g PVP MW 24000, 10 g PEG MW 6000, and 0.5 mL Tween 20 in 1 L of distilled water, pH 8.2), and a 1:10 dilution (w/v) of each sample was used for DAS-ELISA, following the manufacturer's instructions. Positive control was prepared from lyophilized plant tissue infected by GFLV or ArMV (Agritest srl, Valenzano, Italy), and negative control from healthy plant tissue (Agritest srl, Valenzano, Italy), was re-suspended in 2 mL of distilled water and used as positive and negative controls, respectively. Two hours after the addition of the p-nitro-phenylphosphate substrate, the optical densities (O.D.) at 405 nm, using a AMR-100 microplate reader (Hangzhou Allsheng Instruments, China), were measured. The sample was considered positive if its OD 405 value was at least twice the negative control value, as reported in the protocol supplied by Agritest srl.
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!