Spectrostar

Manufactured by BMG Labtech

Sourced in Germany

The SPECTROstar is a multi-mode microplate reader from BMG LABTECH. It is designed to perform various photometric measurements, including absorbance, fluorescence, and luminescence, in microplate formats.

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Lab products found in correlation

Mousewg 6 v2 expression beadchip, by Illumina (2 mentions) Ovation pico wta v2 kit, by Tecan (2 mentions) Biotin il kit, by Tecan (2 mentions) Nanodrop nd 1000, by Thermo Fisher Scientific (2 mentions) Beadarray reader, by Illumina (2 mentions) Bioanalyzer, by Agilent Technologies (2 mentions) Spectrostar nano mars v3, by BMG Labtech (2 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Sybr premix ex taq 2, by Takara Bio (1 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions) Goscript reverse transcription system, by Promega (1 mentions) Spss version 18, by IBM (1 mentions) Astra 6, by Wyatt Technology (1 mentions) Dionex ultimate 3000, by Thermo Fisher Scientific (1 mentions) Dawn heleos 2, by Wyatt Technology (1 mentions) Direct red 80, by Merck Group (1 mentions) Genomestudio, by Illumina (1 mentions) Humanht 12 v4 expression beadchip, by Illumina (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Bioanalyser, by Agilent Technologies (1 mentions)

28 protocols using spectrostar

DPPH Radical Scavenging Assay for Antioxidant Activity

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The antioxidant activity of the extracts was assessed by determining their free radical scavenging activity against 2,2-diphenyl-1-picryldrazyl (DPPH), as described by Brand-Williams [30] . We dissolved 5 mg DPPH in 100 mL ethanol to obtain an absorbance of DPPH at 516 nm between 0.9 and 1.1. Extracts were diluted in ethanol to obtain concentrations of 500 µg/mL. We added a 150 µL aliquot of each extract and 150 µL of 50 µg/mL DPPH in ethanol to the wells of a 96-well microplate. The plate was kept in the dark for 40 minutes, with mixing by gentle rotation every 5 minutes. Absorbance was read at 516 nm with a spectrophotometer (Spectrostar, BMG Labtech, Ortenberg, Germany). Percent inhibition was calculated as follows:
where A is the absorption in the presence of the extract, and A 0 is the absorption of the solution obtained by mixing 150 µL of DPPH solution with 150 µL of ethanol.

Jamaleddine A., Caro P., Bouajila J., Evon P., Haddad J.G., El-Kalamouni C., Hijazi A, & Merah O. (2022). In Vitro Bioactivities of Extracts from Tomato Pomace. Frontiers in bioscience (Landmark edition), 27(9).

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Protein Quantification Using BCA Assay

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The protein concentration was calculated using a bicinchoninic acid (BCA) assay. The colorimetric reaction was measured at 550 nm in a Spectrostar microplate reader (BMG Labtech, Ortenberg, Germany). Samples of total cellular extracts were mixed with a reaction solution [bicinchoninic acid and copper (II) sulfate]. The purple color proportionally increased with the amount of protein. A bovine serum albumin (BSA) curve was used to normalize protein [45 (link)].

Ureña-Vacas I., González-Burgos E., Divakar P.K, & Gómez-Serranillos M.P. (2022). Lichen Extracts from Cetrarioid Clade Provide Neuroprotection against Hydrogen Peroxide-Induced Oxidative Stress. Molecules, 27(19), 6520.

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Apig Dispersion and NEs Stability

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The chemical stabilities of pure Apig dispersion and Apig-loaded NEs were investigated by using a UV-Vis spectrophotometer (SPECTROstar^® Nano BMG LABTECH GmdH, Ortenberg, Germany). The study was done by comparing the sample absorbance with methanolic Apig solution (5% v/v methanol). Absorbance wavelengths at 267 nm and 336 nm were measured in the 0, 1, 2, 3, 4, 6, 12, 18, 24, 30, 36 h.

Chou T.H., Nugroho D.S., Chang J.Y., Cheng Y.S., Liang C.H, & Deng M.J. (2021). Encapsulation and Characterization of Nanoemulsions Based on an Anti-oxidative Polymeric Amphiphile for Topical Apigenin Delivery. Polymers, 13(7), 1016.

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CXCL10 Levels in Periodontal Disease

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The levels of CXCL10 were analyzed in GCF, saliva, and serum using a commercial ELISA kit (Abcam, Cambridge, UK). All samples were thawed at the room temperature. GCF was eluted from the Periopaper by adding 250 µL of PBS and mixing on Vortex. Standards in the commercial ELISA kit were diluted, and test samples were added to wells coated with CXCL10-specific antibodies. All assay procedures were performed according to the manufacturer’s instructions, and the absorbance values were determined by a spectrophotometric ELISA Reader (SpectroStar; BMG LABTECH, Offenburg, Germany). The total amounts of CXCL10 were expressed as picogram/milliliter.
The data were analyzed using the SPSS statistical package software, for Windows version 17.0 (SPSS Inc., Chicago, IL, USA). All the quantitative data are presented as mean and SD. Student’s t-test was carried out to evaluate the significant difference between the different groups. Spearman’s correlation test was used to analyze the relationship between the levels of CXCL10 and clinical periodontal parameters. P-value of ≤0.05 was considered significant.

Aldahlawi S., Youssef A.R, & Shahabuddin S. (2018). Evaluation of chemokine CXCL10 in human gingival crevicular fluid, saliva, and serum as periodontitis biomarker. Journal of Inflammation Research, 11, 389-396.

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Cell Viability Assay for Viral and Drug Effects

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Cells (10,000 cells/96-well) were infected with the virus or treated with drugs or a combination of virus and drugs in 2% FBS/1% P/S DMEM. Cell viability was determined after 3–6 days by the MTS assay (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay; Promega, Southampton, UK) and live cells were detected by a microplate reader (SPECTROstar; BMG Labtech, Ortenberg, Germany). Viral and drug dose–response curves were generated to determine the concentration killing 50% of cells (EC₅₀) using untreated and drug-treated cells as controls. Each data point was generated from triplicate samples and experiments repeated at least three times [17 (link)].

Miao T., Symonds A., Hickman O.J., Wu D., Wang P., Lemoine N., Wang Y., Linardopoulos S, & Halldén G. (2024). Inhibition of Bromodomain Proteins Enhances Oncolytic HAdVC5 Replication and Efficacy in Pancreatic Ductal Adenocarcinoma (PDAC) Models. International Journal of Molecular Sciences, 25(2), 1265.

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Microarray analysis of mouse gene expression

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RNA was assessed for concentration and quality using a SpectroStar (BMG Labtech) and a Bioanalyzer (Agilent Technologies). Microarray experiments were performed at Cambridge Genomic Services, University of Cambridge, using the MouseWG-6v2 Expression BeadChip (Illumina) according to the manufacturer’s instructions. Briefly, Total RNA was amplified using the Ovation Pico WTA V2 kit (NuGEN) and subsequently labeled using the Biotin Il kit (NuGEN). The concentration, purity and integrity of the resulting cRNA were measured using the Nanodrop ND-1000 (Thermo Scientific) and by Bioanalyzer. cRNA was then hybridized to the MouseWG-6 v2 Beadchip overnight followed by washing, staining and scanning using the Bead Array Reader (Illumina). Raw microarray data was preprocessed using the “lumi” bioconductor49 (link) in R. After filtering for significance, data was transformed using the Variance Stabilization Transformation50 (link) from lumi, and normalized using quantile normalization. The data discussed in this study have been deposited in the NCBI Gene Expression Omnibus (GEO; GEO Series accession: GSE73728)

Riedel A., Shorthouse D., Haas L., Hall B.A, & Shields J. (2016). Tumor Induced Stromal Reprogramming Drives Lymph Node Transformation. Nature immunology, 17(9), 1118-1127.

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mRNA Extraction and Quantitative PCR

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mRNA expressed in the liver and the intestine was prepared with TRIzol Reagent (Invitrogen, USA). 50 mg of each samples were homogenised in 1mL Trizol using a T10 basic ULTRA-TURRAX (IKA, Germany) and centrifuged at 12000 g for 10 min. After adding 0.2 mL of chloroform and centrifuging 12000 g for 15 min, only the aqueous phase containing the RNA was transferred to a new tube; RNA precipitation was performed by addition of 0.5 mL of isopropanol. The RNA pellet formed after centrifugation at 7500 g for 10 min was washed with 70% ethanol and resuspended in RNase-free water after removal of the ethanol. The purity and concentration of RNA were measured by SPECTROstar (BMG LABTECH, Germany). 3 μg of cDNA (complementary DNA) were prepared using the GoScript^™ Reverse Transcription System (Promega, USA) and a Verity 96-well thermal cycler (ABI Research, USA). Quantitative real-time PCR was performed by SYBR Premix Ex Taq^™ II (Takara, Japan) with 20 ng of cDNA for each reaction using Step-One Plus real-time PCR System (Applied Biosystems, USA). Data were normalised by glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression of each organ. Specific primers used in this study are:

Park S., Kang J., Choi S., Park H., Hwang E., Kang Y., Kim A., Holzapfel W, & Ji Y. (2018). Cholesterol-lowering effect of Lactobacillus rhamnosus BFE5264 and its influence on the gut microbiome and propionate level in a murine model. PLoS ONE, 13(8), e0203150.

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Microplate Absorbance Measurement Protocol

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All analyzes were performed in 96-well microplates and absorbances were measured in the plate reader (SPECTROstar^® Nano BMG LABTECH GmbH; Offenburg, Germany).

Czubak-Prowizor K., Trelinski J., Stelmach P., Stelmach P., Madon A, & Zbikowska H.M. (2021). Increased Oxidative Stress in Acute Myeloid Leukemia Patients after Red Blood Cell Transfusion, but Not Platelet Transfusion, Results Mainly from the Oxidative/Nitrative Protein Damage: An Exploratory Study. Journal of Clinical Medicine, 10(7), 1349.

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Cytotoxicity Evaluation of SE and PM

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Cytotoxicity Studies The cytotoxicity of SE and PM samples to Caco-2 cells was determined using an MTT assay. The Caco-2 cells were seeded in 96 well plates at a density of 1 × 10 4 cells/mL in 200 µL cDMEM and incubated at 37°C in an atmosphere with 5% CO 2 and 95% relative humidity for 24 h. The old medium was removed, and cells were washed with DMEM. The cells were treated with SE or PM at equivalent concentrations of ORV (12.5, 25, 50, 100, or 200 µM in DMEM) for 24 h. Cells treated with DMEM were used as a control group. After treatment, 20 µL of MTT solution (5 mg/mL in phosphate buffered saline (PBS)) was added to each well and incubated for another 4 h (37°C, 5% CO 2 ). Finally, the medium was removed, and 200 µL of dimethyl sulfoxide (DMSO) was added to dissolve the formazan crystals. The absorbance of formazan was measured at 540 nm by a microplate reader (SPECTROstar, BMG LABTECH, Germany). The experiment was performed in four replicates. The concentrations of SE and PM that produced cell viability that was not significantly different from the control were chosen for further evaluation of cellular transport.

Suzuki Y., Muangnoi C., Thaweesest W., Teerawonganan P., Ratnatilaka Na Bhuket P., Titapiwatanakun V., Yoshimura-Fujii M., Sritularak B., Likhitwitayawuid K., Rojsitthisak P, & Fukami T. (2019). Exploring Novel Cocrystalline Forms of Oxyresveratrol to Enhance Aqueous Solubility and Permeability across a Cell Monolayer. Biological & pharmaceutical bulletin, 42(6).

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Cell Growth Kinetics Monitoring

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Overnight liquid cultures were diluted to 60 cells/μL in 100 μL YPD and incubated at 30°C in a 96 well plate (Cellstar, #655180) with double orbital agitation of 400 rpm using a BMG Labtech SPECTROstar nanoplate reader for 48 hours. When indicated, YPD media was supplemented with MMS (0.005%) and HU (22 mM). Graphs show the mean of 3 biological replicates, error bars show the standard deviation.

Rizzo M., Soisangwan N., Vega-Estevez S., Price R.J., Uyl C., Iracane E., Shaw M., Soetaert J., Selmecki A, & Buscaino A. (2022). Stress combined with loss of the Candida albicans SUMO protease Ulp2 triggers selection of aneuploidy via a two-step process. PLOS Genetics, 18(12), e1010576.

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