The creatinine standard and the urine samples were diluted 100-fold with distilled water and dispensed into a 96-well microplate (AS ONE Co., Ltd., Osaka) at 100 μL each, followed by 50 μL of 1 M NaOH and 50 μL of 1 g/dL trinitrophenol. The plate was left at room temperature (22–26°C) for 20 min, and the absorbance was measured at a wavelength of 490 nm using a microplate reader (MODEL 680XR, Bio-Rad Laboratories, Inc., USA). The samples were used undiluted.
Model 680xr
Manufactured by Bio-Rad
Sourced in United States
The Model 680XR is a microplate reader designed for absorbance-based assays. It can measure the optical density of samples in microplates with 6- to 384-well formats. The device is capable of performing a variety of absorbance-based assays, such as enzyme-linked immunosorbent assays (ELISA), enzymatic reactions, and colorimetric assays.
Automatically generated - may contain errors
Lab products found in correlation
96 well microplate, by As One (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay system, by Promega (1 mentions)
Model infinite 200 pro, by Tecan (1 mentions)
Anaeropack system, by Mitsubishi (1 mentions)
Wst 1 reagent, by Roche (1 mentions)
Gemcitabine, by Fluorochem (1 mentions)
Victor nivo multimode microplate reader, by PerkinElmer (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Cell count reagent sf, by Nacalai Tesque (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Progesterone, by Merck Group (1 mentions)
Estrogen, by Merck Group (1 mentions)
26 protocols using model 680xr
1
Colorimetric Creatinine Quantification
2
Fn Binding Assay for PE_PGRS61 Protein
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The Fn binding assay was performed in corning 96-well microtiter plates (23 (link)). With wash buffer 1- X PBS, a blocking solution 3% (w/v-1) bovine serum albumin (BSA), with dilution buffer, carbonate-bicarbonate buffer [pH 9.6], 1X-PBS and 0.1% BSA. To assay binding of biotinylated Fn to PE_PGRS61 protein the triplicate well of the microtiter plates were coated in carbonate-bicarbonate buffer by using various concentrations of protein in μg as 0.15, 0.20, 0.25, 0.30, and 0.35 respectively in addition to various controls. The microtiter plates were incubated at 37 °C for 1 hr, followed by overnight incubation at 4 °C. After blocking the wells with blocking solution 3% (w/v-1) BSA, wells were incubated with indicated 10 ng and 20 ng concentrations of biotinylated Fn for 1 hr at room temperature on an orbital shaker. After 60 min incubation on a rocking platform, the wells were washed extensively, followed by addition of (1:5000) fold diluted alkaline phosphatase-conjugated streptavidin. Wells were again incubated for 1 hr, washed extensively and developed with 1 mg/ml para-nitrophenyl phosphate substrate for 1 hr in the dark place at room temperature. Plates were read on a microplate reader (Bio-Rad model 680 XR) equipped with 405 nm filter. Data were expressed as the mean absorbance value (A405) of triplicate wells.
3
Evaluating Cell Proliferation with CellTiter 96
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For cell proliferation assays, the CellTiter 96 AQueous One Solution Cell Proliferation Assay System (Promega, Madison, WI, USA) was used according to the manufacturer’s instructions. PC12 cells were plated in each well of a 96-well plate and the indicated concentrations of Wnt3a, sclerostin, XAV939, BMP2, BMP4, and BMP7 was added. After 24-hour incubation in a humidified 5% CO2 atmosphere, PC12 cells were incubated with 20 μL of CellTiter 96 AQueous One Solution reagent per well. The absorbance at 490 nm was measured using a 96-well plate reader (Bio-Rad, Hercules, CA, USA, Model 680 XR).
4
Cationic Lipoplexes for Gene Delivery
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To evaluate the transfection efficiency
of the cationic GAs alone and their formulations to induce gene expression
in HeLa and A549 cells, pCMV·SPORT-β-gal (300 ng) was used
to form lipoplexes at various N/P ratios (1–6).23 (link),39 (link) The cells were seeded in 96-well plates at a density of 5000 cells/well
in DMEM (200 μL) growth medium and penicillin–streptomycin–amphotericin
B (1%) solution. After 18–24 h, the cells were treated with
the diluted lipoplexes in 200 μL plain DMEM per well. After
4 h of incubation of the formulation, the culture media were removed,
cells were washed with PBS (pH 7.4), and complete growth medium (200
μL) was added to each well. Culture media were removed after
a time span of 48 h, and cells were washed with PBS (pH 7.4) and lysed
(using lysis buffer 0.5% Nonidet P-40 in Tris buffer, pH 8.0, 50 μL).
The cells were further treated with ONPG solution (2×, 50 μL),
a substrate for β-galactosidase. The intensity of yellow color
was recorded after 15 min of incubation at 37 °C in an enzyme-linked
immunosorbent assay plate reader (Biorad, Model 680 XR) at 405 nm.
Naked DNA transfected cells were used as the negative control in all
of the experiments.22 (link),24 (link)
of the cationic GAs alone and their formulations to induce gene expression
in HeLa and A549 cells, pCMV·SPORT-β-gal (300 ng) was used
to form lipoplexes at various N/P ratios (1–6).23 (link),39 (link) The cells were seeded in 96-well plates at a density of 5000 cells/well
in DMEM (200 μL) growth medium and penicillin–streptomycin–amphotericin
B (1%) solution. After 18–24 h, the cells were treated with
the diluted lipoplexes in 200 μL plain DMEM per well. After
4 h of incubation of the formulation, the culture media were removed,
cells were washed with PBS (pH 7.4), and complete growth medium (200
μL) was added to each well. Culture media were removed after
a time span of 48 h, and cells were washed with PBS (pH 7.4) and lysed
(using lysis buffer 0.5% Nonidet P-40 in Tris buffer, pH 8.0, 50 μL).
The cells were further treated with ONPG solution (2×, 50 μL),
a substrate for β-galactosidase. The intensity of yellow color
was recorded after 15 min of incubation at 37 °C in an enzyme-linked
immunosorbent assay plate reader (Biorad, Model 680 XR) at 405 nm.
Naked DNA transfected cells were used as the negative control in all
of the experiments.22 (link),24 (link)
5
NAD(H) Quantification in Plasma and CSF
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Total NAD(H) concentrations in plasma and CSF samples were measured spectrophotometrically using a thiazolyl blue microcycling assay established by Bernofsky and Swan (1973) [36] (link), and adapted to a 96 well plate format by Grant and Kapoor (1998) [37] (link). In brief, 125 µL of the reaction mixture containing 120 mM bicine (pH 7.8), 0.5 mM MTT, 2 mM PMS, 0.6 M ethanol and alcohol dehydrogenase (300 units/mL) was added to either 6 µL of plasma or 20 µL of CSF. Following a 10 minute incubation at 37 degrees Celsius, the concentration of total NAD(H) was measured, using a Model 680XR microplate reader (BioRad, Hercules), as the change in absorbance at 570 nm.
6
MTT Assay for Cell Growth Analysis
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For assessment of cell growth of EPC2 and BAR-T cells, we performed a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay77 (link). We seeded 1000 cells per well in 96 well plates for each of the three wild-type and three HOXA13 knock-out cell lines. Per condition at least 2 wells were used. On days one, three, five, and seven 10 µl MTT at 5 µg/ml was added and incubated for three hours, the medium was removed, and the precipitate was dissolved in 100 µl DMSO, which was incubated for five minutes under continuous shaking. For BAR-T cells, absorption was measured in a BioRad microplate reader Model 680 XR at 490 and 595 nm, the average absorption was used to process the data. For EPC2 cells it was measured with Tecan microplate reader Model Infinite 200 pro at 565 nm with reference wavelength 670 nm. The experiment was repeated three times and a two sided Student’s t-test was used to test for statistical significance.
7
Quantifying Streptococcus mutans Biofilm
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Overnight culture of S. mutans strain UA159 (ATCC 700610) in BHI medium was inoculated into 200 μL of CDM per well (3.0 × 106 CFU) of the flat-bottomed 96-well microtiter plate (Corning, 3595). After 18 h of standing incubation at 37 °C in 5% CO2 under anaerobic condition in an AnaeroPack system (Mitsubishi Gas Chemical, Tokyo, Japan), the cultured medium was removed, and adherent bacteria were stained with 200 μL of 0.005% crystal violet for 30 min. The wells were washed twice with 300 μL of distilled water and then air dried. The dye was extracted into 200 μL of 60% ethanol, and the biofilm mass was estimated by using a microplate reader (Bio-Rad, Model 680XR) (Bio-Rad, Hercules, CA) to measure the absorbance at 595 nm. The bacterial growth was determined by measuring the turbidity (absorbance at 595 nm) of parallel wells.
For the count of viable cells shown in Fig.2B , part of the culture was serially diluted and spread onto the BHI plate, and the plates were incubated overnight at 37 °C in 5% CO2 under anaerobic condition.
For the count of viable cells shown in Fig.
8
Hypoxia-Induced Drug Cytotoxicity Assay
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Cells were seeded at a density of 2 × 103 cells/well in 96‐well plates and pre‐incubated for 12 h in normoxia. TS‐1 (Taiho, Tokyo, Japan) and gemcitabine (Fluorochem, Hadfield, UK) were added to the medium as a dilution series of 0–1000 μM and 0–100 nM, respectively, before incubation in 21%, 1%, or 0.1% O2. After incubation for 72 h, 10 μL WST‐1 reagent (Roche Diagnostics) was added to each well. After a 3‐h incubation in normoxia, the optical density (450 nm) of each well was measured with the reference optical density (750 nm) using a microplate reader Model 680XR (Bio‐Rad, Hercules, CA, USA).
9
Biofilm Quantification Assay with Antibiotics
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A microtiter plate assay was performed in 96-well polystyrene microtiter plates (Orange Scientific, Belgium), to quantitatively determine the biofilm formation at different time intervals (72, 120, and 172 hours) in the presence and absence of antibiotics (0.1 µg/mL) and plant crude extracts (2 mg/mL) (14 (link), 15 (link)). After the processing of the assay, the OD was measured at 578 nm, with the help of a microplate reader (BIO-RAD Model 680 XR). Those microtiter wells containing un-inoculated LB broth were considered to be the negative controls, while the inoculated wells were positive controls.
10
Quantifying Melanin in B16-F1 Cells
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The Melanin assays were performed essentially as described previously [31 (link)]. In brief, the B16-F1 cells (WT and KO) were solubilized in 50 mM HEPES-KOH, pH 7.2, 150 mM NaCl, and 1% Triton X-100. The pigment was then pelleted by centrifugation at 20,400× g for 10 min at 4 °C, and the pellet was dissolved in 2N NaOH/20% DMSO for 30 min at 100 °C. Melanin content was measured as optical density at 490 nm with a microplate reader (model 680XR, Bio-Rad; Figure 2 E) or Victor Nivo Multimode microplate reader (PerkinElmer, Waltham, MA, USA; Figure 5 D) and normalized to total protein content.
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