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Stat Fax 2100 Microplate Reader

Manufactured by Awareness Technology
Sourced in United States, China

The Stat Fax 2100 Microplate Reader is a compact and versatile instrument designed for absorbance-based assays. It can read microplates with up to 96 wells, utilizing a single-beam optical system with a tungsten-halogen light source. The reader is capable of performing absorbance measurements at a wide range of wavelengths, enabling a variety of applications in life science research and clinical diagnostics.

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19 protocols using Stat Fax 2100 Microplate Reader

To study 5-Azacytidine cytotoxicity cells were seeded onto 96-well plates (3 × 103 cells per well) and incubated overnight. Then, the medium was replaced with a fresh one containing 0.5, 1, 2, 3–10, 20, 30–90 or 100 μM 5-Azacytidine. Cells were incubated at 37˚C in the atmosphere of 5% CO2 for 72 h. The medium with 5-Azacytidine was changed every 24 h. Cell viability was evaluated using MTT assay [26 (link)]. The optical density was measured on a StatFax-2100 Microplate reader (Awareness Technology, USA) at a wavelength of λ = 545 nm. An example of a cell survival curve (for HeLa cells) is shown in S2 Fig.
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The study of cell proliferation was carried out through the crystal-violet assay. For this purpose, two groups of 5000 MCF-7 and HCC1937 cells were inoculated in 96-well culture plates and were left to grow during 24 h (group 1) and 48 h (group 2). Afterwards, each group was washed every 24 h, during 96 h, with PBS and fresh culture medium (supplemented with 5% SFB) with leptin stimuli, which was added (Recom Hu Leptin Active, Sino Biological, Thermo Fisher Scientific, USA) in concentrations of 0 ng/mL (negative control), 10 ng/mL, 100 ng/mL, and 1000 ng/mL [27 (link)]. The decrease of SFB was done to reduce the growth factors that could interfere with the action of leptin [40 (link)].
Cell proliferation in each cell line was determined in triplicate for every concentration evaluated. After 24 h of stimulation, the cells were fixed with 1.1% glutaraldehyde (Merck, Germany) during 15 minutes and then PBS was added until the 96 h of stimulation with leptin was concluded for each group. At the end of stimulation, the fixed cells were stained with crystal-violet for 15 minutes, the plate was washed with water, and 10% acetic acid was placed in each well. Immediately after, the plate was read at an excitation wavelength of 490 nm and an emission wavelength of 630 nm in a microplate reader (Stat Fax 2100 Microplate Reader, Awareness Technology Inc.)
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Human umbilical vein endothelial cells (20000 cells/well) were cultured according to the treatment above after which 20 mL of the stock solution of MTT (5 mg/mL) was added to each well for 4 h. Finally, the incubation medium was removed, and the formazan crystals were dissolved in 150 μL of DMSO. The MTT reduction was measured as the absorbance at 490 nm using a StatFax-2100 Microplate Reader (Awareness Technology Inc., Palm City, FL). The background absorbance of the control wells was subtracted. The analysis was performed in triplicate. Viability (%) = OD (experiment group)/OD (control) × 100%.
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ELISA kits for human IL-17 (sensitivity: <1 pg/ml), IL-5 (sensitivity: <1 pg/ml), IL-9 (sensitivity: 0.5 pg/mL) and TGF-β (sensitivity: <1 pg/ml) were purchased from Peprotech (Rocky Hill, NJ, USA) and ELISA kits for human IFN-γ (sensitivity: ≤ 2 pg/mL) and IL-8 (sensitivity. 2.0 pg/mL) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Analyses were performed according to the manufacturers' instruction for each ELISA kit. Absorbance was read by StatFax-2100 microplate reader (Awareness Technology Inc., USA).
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In order to determine the cytotoxicity of ATF3, MTT assay was performed. Briefly, cells were seeded into 96-well plates at the density of 7000 cells per well in 200 µL DMEM medium and incubated at 37‎°C‎ and ‎5% CO2. Once Ca Ski cells were grown to the confluence of 70%, they were incubated with calcium phosphate-precipitated pCMV6-ATF3 and mock plasmid with concentrations ranging between 0.1 and 1 µg according to the above-mentioned transfection protocol. Cell viability was determined 24, 48 and 72 h post-transfection by adding fresh medium containing 20 µL MTT solution (5 mg/mL) (Sigma-Aldrich), followed by a four-hour incubation for crystal formation. Next, ‎150 µL of DMSO (Sigma-Aldrich) was added‎ to each well. Finally, the absorbance was measured at the wavelength of 570 nm with a Stat Fax 2100 microplate reader (Awareness Technology Inc., Palm City, FL, USA). ‎The MTT assay was performed in triplicate.
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Waist and hip circumferences, body weight, and height were measured in all subjects as anthropometric variables. The body mass index (BMI) and waist–hip ratio (WHR) were calculated as follows:
BMI = body mass/ (height)2[kg/m2],
WHR = waist circumference (cm)/hip circumference (cm).
For measurement of the serum levels of follicle-stimulating hormone (FSH; Monobind kit, USA), luteinizing hormone (LH; Monobind kit, USA), testosterone (Monobind kit, USA), estrogen (Monobind kit, USA), insulin (Monobind kit, USA), and TNF-α (eBioscience, Austria), the enzyme-linked immunosorbent assay (ELISA) method was used according to the manufacturer’s recommendations. Color intensities at the final step were recorded using an ELISA reader (Stat Fax-2100 microplate reader, Awareness Technology, USA). The biochemical parameters including fasting blood glucose (Pars azmoon, Iran), triglyceride (Pars azmoon, Iran), total cholesterol (Pars azmoon, Iran), low-density lipoprotein (LDL; Pars azmoon, Iran), and high-density lipoprotein (HDL; Pars azmoon, Iran) were measured using a BT3000 autoanalyzer (Biotechnica Instruments, USA). Homeostasis model assessment (HOMA) as an insulin resistance index was computed using the following formula: HOMA = fasting glucose (mg/dl) × fasting insulin (mU/ml)/405.
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Positive results observed by LAT were further confirmed using a commercial Toxoplasma IgG ELISA kit (Atlas Medical Company, UK). ELISA was performed following the manufacturer’s instructions. Anti-human enzyme conjugate was used for human sera, and anti-multispecies horseradish peroxidase conjugate was used (IDvet Innovative Diagnostics, Grabels, France) for sheep samples. The optical density was measured at 450 nm using Stat Fax 2100 Microplate Reader (Awareness Technology, Inc., FL, USA).
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An enzyme-linked immunosorbent assay (ELISA) kit (Sino-UK Institute of Biological Technology, Beijing, China) with a STAT FAX 2100 Microplate Reader (Awareness Technology Inc., Palm City, FL, USA) was used for measurements of estradiol (E2), free testosterone (FT), sex hormone-binding globulin (SHBG), luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), and serum fasting insulin (FINS) concentrations. All indices were tested during the diestrus stage after 12–14 h of fasting (21 (link)).
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The total protein concentration in human skim milk samples was determined by bicinchoninic methods with the Bicinchoninic Acid Protein Assay Kit (Sigma, St. Louis, MO, USA) (37 (link)). For testing 25 μL of 12.5- and 25-fold diluted in TRIS-buffered saline (TBS, pH 7.5), skim milk samples and bovine albumin, as a standard from 0.2 to 1.0 mg/mL (SERVA, Heidelberg, German), were prepared and transferred to the wells of microtiter plates, and then 200 μL of freshly prepared bicinchoninic acid working reagent (solution of bicinchoninic acid and copper (II) sulfate in the ratio 1:50, respectively) was added. The plates were incubated at 37°C for 35 min and the absorbance was measured at 560 nm in a Stat Fax 2100 Microplate Reader (Awareness Technology Inc., Palm City, FL, USA). All skim milk samples were analyzed in duplicate.
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10

Dose-Dependent Tamoxifen Effects on Breast Cancer Cell Lines

Proliferation of 5,000 cells of the cell lines MCF-7, MDA-MB 231 and HCC1937 (n = 3) was carried out in 96-well plates for 48 hours in RPMI-1640 medium (10% SFB). Subsequently they were stimulated for 48 hours with the following concentrations of tamoxifen: 0.002 μM, 0.02 μM, 0.2 μM, 2 μM, 20 μM and 200 μM, changing the stimulus every 24 hours.
At the end of the 48 hours of stimulation, the cells were fixed with 4% formaldehyde for 15 minutes. The fixed cells were washed with PBS and stained with crystal violet (0.02%) for 15 minutes, to remove the residual dye, 3 washes of 250 μl of water were carried out and finally 250 μl of 10% acetic acid was added to each well. The plate was read at an excitation wavelength of 490 nm and an emission length of 630 nm in a microplate reader (Stat Fax 2100 Microplate Reader, Awareness Technology Inc.).
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