The total content of phenolic compounds in peas were determined according to the colorimetric method described with some modifications (Kan et al., 2017 (link)). Firstly, 20 µL gallic acid standards (0.025–1.6 mg/mL) or samples were mixed with 20 µL Folin-Ciocalteu reagent (0.2 M) in a 96 well plate and reacted for 30 s at 37 ℃ in a temperature incubator (SANYO Electric Co., ltd., Osaka, Japan), and then 60 µL saturated sodium carbonate (Na2CO3, 0.1 g/mL) solution was added for reacting 15 min. The absorbance was measured at 764 nm using a ELX800 microplate reader (Bio-Tek Instruments, Inc., Winooski, VT, USA). In addition, the total flavonoids content (TFC) was estimated using the colorimetric assay according to previous report with some modifications (Chang, Yang, Wen, & Chern, 2002 (link)). A volume of 5 µL rutin standard or sample solution was mixed with 10 µL sodium nitrite (0.05 g/mL) solution and 40 µL distilled water in 96 microporous plate for reacting 5 min at 37 ℃ and then 10 µL aluminum chloride (0.1 g/mL) was added to react for 6 min. Finally, 30 µL sodium hydroxide (0.1 M) solution was added and reacting for 15 min. The absorbance of the mixture was then measured at 510 nm using a ELX800 microplate reader (Bio-Tek Instruments, Inc., Winooski, VT, USA). All the samples were analyzed in triplicate.
Elx800 microplate reader
Manufactured by Agilent Technologies
Sourced in United States, China, United Kingdom, France, Canada, Japan
The ELx800 microplate reader is a compact and versatile instrument designed for various absorbance-based assays. It can measure the optical density of samples within a microplate format. The ELx800 is capable of performing rapid, high-throughput measurements and provides accurate quantitative data.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (7 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (7 mentions)
Cck 8, by Dojindo Laboratories (5 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (5 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Cell counting kit 8 (cck8), by Beyotime (4 mentions)
96 well plate, by Corning (4 mentions)
2 n h2so4, by Merck Group (3 mentions)
Cck 8 assay, by Beyotime (3 mentions)
Cell counting kit 8 (cck8), by Abcam (3 mentions)
Cck 8 reagent, by Dojindo Laboratories (3 mentions)
Cck 8 reagent, by Merck Group (3 mentions)
Tacs mtt cell proliferation and viability assay kit, by Agilent Technologies (2 mentions)
Tmb solution, by Merck Group (2 mentions)
Innoscan 300 microarray scanner, by Innopsys (2 mentions)
Quantibody mouse cytokine array 1 kit, by RayBiotech (2 mentions)
Raybio mouse ifn γ kit, by RayBiotech (2 mentions)
Mtt solution, by Merck Group (2 mentions)
Cell death detection elisaplus kit, by Roche (2 mentions)
Cck 8, by Beyotime (2 mentions)
372 protocols using elx800 microplate reader
1
Quantifying Phenolics and Flavonoids in Peas
2
Quantification of Phenolic and Flavonoid Content
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Folin–Ciocalteu technique, with a little modification, was used to perform the analysis for the Total Phenolic Content (Antony et al., 2013 (link)). The NPs samples (4 mg/mL) were moved into 96 well plates, and 90 µL of Folin–Ciocalteu reagent was added to each well and reaction mixture was nurtured for five minutes before sodium carbonate was add alsoed to the plate. The absorbance of the reaction mixture after one hour incubation was measured at 630 nm using a microplate reader (Biotek USA, microplate reader Elx 800). The sample of gallic acid that was utilized was considered as the standard. The results obtained were presented as per milligrams of the gallic acid equivalents (GAE/mg).
The Total flavonoid content was also calculated using the aluminum chloride method (AlCl3) (Paulkumar et al., 2014 ). Each well of the plate containing 10 % AlCl3 solution, purified water, and potassium acetate was filled with 20 µL of NP sample (4 mg/mL). After 30 min of incubation at room temperature, the plate was analyzed. The microplate reader recorded the sample absorbance at 630 nm. Two different substances, quercetin and DMSO, served as positive and negative controls, respectively. Three replicate reactions were used to calculate the quercetin equivalent per milligrams (QE/mg) in the final extracts.
The Total flavonoid content was also calculated using the aluminum chloride method (AlCl3) (Paulkumar et al., 2014 ). Each well of the plate containing 10 % AlCl3 solution, purified water, and potassium acetate was filled with 20 µL of NP sample (4 mg/mL). After 30 min of incubation at room temperature, the plate was analyzed. The microplate reader recorded the sample absorbance at 630 nm. Two different substances, quercetin and DMSO, served as positive and negative controls, respectively. Three replicate reactions were used to calculate the quercetin equivalent per milligrams (QE/mg) in the final extracts.
3
Cell Viability Assay of Hepatocellular Carcinoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability assays was conducted according to the method described in previous study [25 (link)]. SNU-449 and Hep-3B cells (5 × 103 per well) were seeded into a 96-well plate in a 5% CO2 incubator overnight. SNU-449 and Hep-3B cells were respectively exposed to various doses of SK (0, 0.25, 0.5, 1, 2, 4 and 8 µM) (0, 0.5, 1, 2, 4, 8 and 16 µM) for 24 h, 48 h, 72 h and 96 h or transfected with PYCR1 siRNA for 48 h. Then, the cell counting kit (CCK)-8 solution (10 μL) (Beyotime, Shanghai, China) was added to each well. Absorbance at 450 nm was measured using a Microplate Reader (ELx800, BioTek, USA).
4
Antioxidants and Oxidative Stress Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The activity of SOD (U/gram tissue) and GPX (nmol/min/gram tissue) was determined by an ELISA reader (Absorbance Microplate Reader ELx 800 TM BioTek®, USA) using the commercially available kits (Cayman Chemical Company, Ann Arbor, MI, USA). MDA (Cayman Chemical Company) and total ROS (eBioscience, San Diego, CA, USA) were also measured according to the manufacturer's instructions.
5
Anti-HBV Activity Evaluation of Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The anti-HBV activities of compounds 1 and 2 on inhibition of viral HBsAg production in cultured HepG2.2.2.15 cells were evaluated as described elsewhere (Arbab et al., 2017 , Parvez et al., 2019 (link)). Briefly, cells were treated with the fresh media containing 150 μl of various doses of 1 and 2 including controls for 3 days, and assayed for HBsAg expression (Monolisa HBsAg ULTRA, BioRad, USA) as per the kit’s manual. The absorbance was recorded (Microplate Reader ELx800; BioTek, USA) and data was analyzed to estimate inhibition of HBsAg in relation to untreated control.
6
Antiproliferative Activity Evaluation
7
RGC-5 TNF-α Concentration Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A RGC-5 TNF-α concentration assay was performed using a commercial TNF-α ELISA kit (Invitrogen, CA, USA). The detailed processes were conducted according to the manual included in the kit. Positive control: the antibody tested in the kit was replaced by mouse TNF-α (provided in assay, concentration: 720 ng/L). Equal quantities of protein (80 μg) were analyzed in every tested well and the measurements were carried out by Bio-tek microplate-reader (ELx800, IL, USA). The percentage of TNF-α concentration in normal control group was set as 100%. All experiments were repeated at least twice.
8
Antioxidant Activity Assay of Cellobiose Dehydrogenase
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antioxidant activity of cellobiose dehydrogenase was determined using the DPPH equivalent, according to an adapted colorimetric procedure described by Paduch et al. [46 ] with slight modification. This method is based on the ability of 1,1-diphenyl-2-picrylhydrazyl (DPPH), a stable free radical, to decolorize in the presence of antioxidants. The tested compound (0.1 ml) at concentrations ranging from 6.25 to 800 μg/ml was added to 0.1 ml of DPPH. solution (0.2 mg/ml in ethanol). The absorbance was measured spectrophotometrically at 515 nm using a Microplate Reader Elx800 (BioTek, Winooski, VT, USA) after 15 min (the time required to achieve the reaction plateau) of incubation at room temperature.
The capability of scavenging DPPH. radicals was calculated by the following formula: where A0 means the absorbance of the control sample and A1 means the absorbance of the standards or tested compounds.
The capability of scavenging DPPH. radicals was calculated by the following formula: where A0 means the absorbance of the control sample and A1 means the absorbance of the standards or tested compounds.
9
Cell Proliferation Assay of Seaweed Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The S. monoica derived compounds: NSQ, SND, AES, NDX, ABN, BMN were tested for their cell proliferative or growth stimulatory activities in cultured HUVEC cells. Cells were treated with the different doses of the compounds, including untreated control (0.5% DMSO) for 3 days. MTT assay (TACS MTT Cell Proliferation Assay Kit, Tervigen, USA) was performed as per the kit’s manual. Briefly, the MTT reagent (10 μl/well) was added and incubated in dark for about 4 h at room temperature (RT) until purple color appeared. Further the detergent solution (100 μl/well) was added and the cells were incubated for another 1.5 h at 37 °C. The optical density (OD; λ = 570) was measured (Microplate reader ELx800; BioTek, USA) and data was analyzed by non-linear regression (Excel software 2010; Microsoft, USA) to determine the cell proliferation in relation to the untreated control [%Cell proliferation = (ODsample − ODblank/ODcontrol − ODblank) x100]. All samples were tested in triplicate and repeated.
10
Caspase-3/7 Activation Assay in HUVEC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Based on the promising anti-oxidative and anti-apoptotic activities, an optimal dose (25 μg/ml) of the compounds was tested for cellular caspase-3/7 activation in HUVEC cells in 96-well plates (Set-I: DCF treated and Set-II: MGO treated). Day 3 post-incubation, cellular caspase expressions were measured (Apo-ONE-cas3/7 assay kit; Promega, USA) as per the kit’s manual., Briefly, 100 μl of caspase-3/7 reagent was added to each well, mixed by gentle rocking and incubated in dark for ~ 6 h at RT. Caspase reagent plus culture medium served as blank while reagent plus DMSO treated cells acted as negative control. The OD was measured (Microplate reader ELx800; BioTek, USA) and data was analyzed. All samples were tested in triplicate and repeated.
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!