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Biotek elx800

Manufactured by Agilent Technologies
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The BioTek ELx800 is a microplate absorbance reader designed for a variety of applications in life science research and clinical diagnostics. It features a high-performance monochromator-based optical system and can measure absorbance across a wide wavelength range. The instrument is capable of performing absorbance-based assays, including enzyme-linked immunosorbent assays (ELISAs), cell-based assays, and colorimetric assays.

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Lab products found in correlation

Mushroom tyrosinase solution, by Merck Group (4 mentions) L dopa, by Merck Group (4 mentions) Porcine pancreatic elastase, by Merck Group (4 mentions) N succ ala ala ala p nitroanilide aaavpn, by Merck Group (4 mentions) Cck 8, by Agilent Technologies (3 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (3 mentions) Falgpa, by Agilent Technologies (3 mentions) Cck 8, by Dojindo Laboratories (2 mentions) Mtt reagent, by Merck Group (2 mentions) Falgpa, by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) 96 well plate, by Corning (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Cell proliferation reagent wst 1, by Roche (1 mentions) Facstar cytofluorometer, by BD (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Dingguo (1 mentions) Kbm 2 medium, by Lonza (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)

79 protocols using biotek elx800

1

IGF-2 Enhances HemSCs Proliferation

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Logarithmic growth phase HemSCs were seeded into 96-well tissue culture plates (Corning Incorporated) at an initial density of 2×103. After serum starvation for 24 h, IGF-2 (cat. no. 100-12-50UG; PeproTech, Inc., Rocky Hill, NJ, USA) at 0, 10, 20, 100, and 200 ng/ml, respectively, was added to EBM-2 containing 5% FBS in the experimental groups. After 72 h in the culture, CCK-8 reagents (Dojindo Molecular Technologies, Inc., Kumamoto, Japan) were added to disparate wells under different treatments. The absorbency was read at 490 nm using a microplate reader (Biotek ELx 800; BioTek Instruments, Inc., Winooski, VT, USA).
To examine the growth kinetics of IGF-2-treated HemSCs, HemSCs were seeded into 96-well plates (Corning Incorporated) at a density of 1.5×103 cells/well. After serum starvation, the cells were treated with 100 ng/ml IGF-2 containing 5% FBS. CCK-8 was added at 0, 1, 3, 5, and 7 days after culture and incubated at 37°C for 4 h. The results of the OD values at 490 nm were tested by a microplate reader (Biotek ELx 800; BioTek Instruments, Inc.).
Zhang K., Wang F., Huang J., Lou Y., Xie J., Li H., Cao D, & Huang X. (2018). Insulin-like growth factor 2 promotes the adipogenesis of hemangioma-derived stem cells. Experimental and Therapeutic Medicine, 17(3), 1663-1669.
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2

Tyrosinase Inhibition Assay Using L-DOPA

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The previously described method of [49 (link)], using L-DOPA (5 mM; Sigma Aldrich) was used for performing tyrosinase assay. L-DOPA diphenolase substrate was mixed with 10 μL of test sample along with sodium phosphate buffer (50 mM, pH 6.8). The final volume of the reaction mixture was raised to 200 μL by adding 0.2 mg/mL of mushroom tyrosinase solution (Sigma Aldrich). The extraction solvent replacing the tested sample was used as control. Microplate reader (BioTek ELX800; BioTek Instruments) was used to trace the reaction processes at 475 nm. Relative to corresponding control tyrosinase effect was expressed as percent inhibition.
Jan H., Usman H., Shah M., Zaman G., Mushtaq S., Drouet S., Hano C, & Abbasi B.H. (2021). Phytochemical analysis and versatile in vitro evaluation of antimicrobial, cytotoxic and enzyme inhibition potential of different extracts of traditionally used Aquilegia pubiflora Wall. Ex Royle. BMC Complementary Medicine and Therapies, 21, 165.
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3

Quantification of Secreted HBD-3 Levels

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The culture supernatants were collected, and the amount of secreted HBD-3 was detected using an HBD-3 ELISA kit (cat no. JL19214; Shanghai Jiang Lai Biotechnology Co., Ltd., Shanghai, China) according to the manufacturer's instructions. Absorbance at 490 nm was determined with a microplate reader (BioTek ELx800; BioTek Instruments, Inc., Winooski, VT, USA). Each sample was analyzed in triplicate.
Zhu M., Miao B., Zhu J., Wang H, & Zhou Z. (2017). Transplantation of periodontal ligament cell sheets expressing human β-defensin-3 promotes anti-inflammation in a canine model of periodontitis. Molecular Medicine Reports, 16(5), 7459-7467.
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4

Cell Viability Assay using CCK-8

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Cell viability was checked using the CCK-8 according to the manufacturer’s instructions (Dojindo, Kumamoto, Japan). All transfected cells (2 × 103/100 μL cells per well) were incubated in 96-well plates for 24, 48, 72 and 96 h. The CCK-8 (10 μL) was added to each well and the plates were incubated for 2 h at 37 °C in a dark room, and then the absorbance was determined at 450-nm wavelength (OD450) using a microplate reader (BioTek Elx800; BioTek Instruments, Winooski, VT, USA).
Li D., Hu J., Li S., Zhou C., Feng M., Li L., Gao Y., Chen X., Wu X., Cao Y., Hao B, & Chen L. (2023). LINC01393, a Novel Long Non-Coding RNA, Promotes the Cell Proliferation, Migration and Invasion through MiR-128-3p/NUSAP1 Axis in Glioblastoma. International Journal of Molecular Sciences, 24(6), 5878.
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5

Porcine Elastase Inhibition Assay

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Elastase assay was performed by using porcine pancreatic elastase (Sigma Aldrich) and its activity was determined with spectrophotometer by making use of N-Succ-Ala-Ala-Ala-p-nitroanilide (AAAVPN; Sigma Aldrich) as a substrate and following p-nitroaniline release at 410 nm using a microplate reader (BioTek ELX800; BioTek Instruments) by adopting the method of Wittenauer et al. (2015) [78 (link)]. Triplicated measurements were used and the anti-elastase activity was expressed as a % of inhibition relative to the corresponding control (adding same volume of extraction solvent) for every extract.
Abbasi B.H., Siddiquah A., Tungmunnithum D., Bose S., Younas M., Garros L., Drouet S., Giglioli-Guivarc’h N, & Hano C. (2019). Isodon rugosus (Wall. ex Benth.) Codd In Vitro Cultures: Establishment, Phytochemical Characterization and In Vitro Antioxidant and Anti-Aging Activities. International Journal of Molecular Sciences, 20(2), 452.
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6

Serum Biochemical Profile Analysis

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Serum samples were incubated on ice for 10 min and centrifuged for 10 min at 3000× g. The contents or activities of total protein (TP), albumin (ALB), uric acid (UA), blood urea nitrogen (BUN), calcium (Ca), phosphorus (P), alkaline phosphatase (AKP), glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) in serum were determined by kits, which were purchased from Nanjing Jiancheng Institute of Biological Engineering. The measuring operation was carried out according to the instructions. A microplate reader (Biotek ELX800; Biotek Instruments, Inc., Winooski, VT, USA) was used in the determination.
Zhou J., Yao J., Bai L., Sun C, & Lu J. (2021). Effects of Dietary Supplementation of gEGF on the Growth Performance and Immunity of Broilers. Animals : an Open Access Journal from MDPI, 11(5), 1394.
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7

Cytotoxicity of Ganoderma Triterpenoids on MCF7 Breast Cancer

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The cytotoxicity of Ganoderma triterpenoids against a breast cancer cell line MCF7 (Riken Cell Bank, Ibaraki, Japan) was measured with the Cell Proliferation Reagent WST-1 (Roche, Basel, Switzerland). Briefly, confluent MCF7 cells in 96-well plates were treated with test samples for 72 h. WST-1 reagent (10 μL) was then added to each well. After 2 h incubation, the formazan dye was quantified by absorbance at 450 nm in a microplate reader (Biotek-ELX800, BioTek, Winooski, VT, USA).
Zhu Q., Bang T.H., Ohnuki K., Sawai T., Sawai K, & Shimizu K. (2015). Inhibition of neuraminidase by Ganoderma triterpenoids and implications for neuraminidase inhibitor design. Scientific Reports, 5, 13194.
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8

Apoptosis Assay in Mixed Cell Populations

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Mixed cell populations containing 10% CBRH7919tk and 90% CBRH7919WT cells were seeded in quadruplicate in 96-well plates at 3 × 103 cells per well. When the cells had adhered on the second day, they were exposed to resveratrol (10 µM or 20 µM), GCV (15.7 µM) and DMSO (negative control) for 24 h, and GCV (15.7 μM) was added to the culture medium in one group for 24 h more. Cell viability was assessed using the MTT assay. Absorbance values were determined at 490 nm with a microplate reader (Bio-Tek ELx800; Bio-Tek Instruments Inc., Winooski, VT, USA). To determine the rate of apoptosis, the drug-treated cells were trypsinized, fixed in 70% ethanol at 4°C and stained with 40 mg/mL propidium iodide for 30 min at 37°C. Ten thousand cells from each group were analyzed with a FACStar cytofluorometer (BD Biosciences) that was equipped with an argon-ion laser (488 nm).
Xiao J., Wang X., Wu Y., Zhao Q., Liu X., Zhang G., Zhao Z., Ning Y., Wang K., Tan Y, & Du B. (2018). Synergistic effect of resveratrol and HSV-TK/GCV therapy on murine hepatoma cells. Cancer Biology & Therapy, 20(2), 183-191.
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9

Evaluating Antioxidant Activity in Fish Oils

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The CAA assay previously adapted by de la Fuente et al. [16 (link)] for salmon head oil was used to evaluate the antioxidant activity of the fish oils at 500–2000 µg/mL in DMSO:H2O (1:1 v/v). This assay employs the cell-permeable fluorescent probe dye DCFH-DA, which can be diffused into RAW 264.7 cells and deacetylated by cellular esterases into the nonfluorescent polar derivative DCFH, which switches rapidly to the highly fluorescent DCF when oxidized by intracellular ROS and other peroxides. The fluorescence intensity was measured in a microplate reader (Biotek ELX800, Bio-Tek Instruments, Inc., Winooski, VT, USA) with fluorescence filters for an excitation wavelength of 485 nm and an emission wavelength of 535 nm at 37 °C. AAPH was used as a free radical generator, and quercetin at 0.3 µg/mL was used as a positive control. Results were expressed as the percentage of oxidation inhibition.
Pinela J., de la Fuente B., Rodrigues M., Pires T.C., Mandim F., Almeida A., Dias M.I., Caleja C, & Barros L. (2022). Upcycling Fish By-Products into Bioactive Fish Oil: The Suitability of Microwave-Assisted Extraction. Biomolecules, 13(1), 1.
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10

Cell Viability Assay with siRNA

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HCC-1937, MX-1 and MCF-7 cells in the logarithmic phase were simultaneously digested with trypsin, re-suspended in complete culture medium and arranged in four groups: The siRNA-5 group, the siRNA-3 group, the NC group and HCC-1937 or MX-1 or MCF-7 cell group. Four groups of cells were separately seeded on 2×103 cells/well in three 96-well plates, wherein every cell group was arranged in 6-wells, incubated in 5% CO2 for 24 h at 37°C and mixed with the small interfering RNA. Following cultivation of the four cell groups for 24, 48 and 72 h, 10 µl of MTT (5 mg/ml) (Beijing Dingguo Biotechnology, Beijing, China) was separately added to each well. The medium was discarded 4 h later and 100 µl dimethyl sulfoxide (DMSO; Shanghai Pharmaceutical Group Co., Ltd., Shanghai, China) was added to each well to stop the reaction. After vortexing for 10 min at room temperature, optical density was measured at 490 nm using a microplate reader (Biotek ELx800; BioTek Instruments, Winooski, VT, USA).
Zhu L., Zhou W., Zhu X., Xiang S., Wang S., Peng Y., Lu B., Tang P., Chen Q., Wu M., Peng X., Chen Z., Sun Z., Yang K., Xiang M, & Yu D. (2018). Inhibitor of apoptosis protein-like protein-2: A novel growth accelerator for breast cancer cells. Oncology Reports, 40(4), 2047-2055.
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