Multiscan spectrum microplate reader

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Multiscan Spectrum microplate reader is a versatile laboratory instrument designed for a wide range of absorbance-based assays. It is capable of measuring absorbance across a broad wavelength spectrum, enabling researchers to perform various types of spectrophotometric analyses on samples in microplates.

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Htf 1, by BD (1 mentions)

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Microplate reader (4490 citations) Multiscan Spectrum (58 citations) Multiscan microplate reader (7 citations) Multiscan spectrum reader (2 citations)

6 protocols using multiscan spectrum microplate reader

MC Proliferation Assay Protocol

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After transfection, MCs were inoculated in 96‐well plates at 3000 cells per well and cultured at 37°C with 5% CO₂. The proliferation rates of MCs were detected using a cell proliferation and toxicity detection kit (CCK‐8, cat. no. MA0218‐500T, Meilunbio). The absorbance in each group was measured at 450 nm using a Multiscan Spectrum microplate reader (Thermo Fisher, Waltham, MA, USA). This experiment was performed three times.

Li Y., Wang S., Ning J., Mao X., Ge K, & Zhang R. (2023). The effects of miRNA‐27a‐3p on human epidermal melanocytes. Skin Research and Technology, 29(5), e13345.

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Cell Lysis and L-DOPA Assay

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Transfected cells were added 90 µL concentration 1% TritonX-100 solution in each hole and placed in −80 °C refrigerator frozen stored for 30 min and made cell lysis completely. Then, 10 µL 0.1% l-DOPA solution was added in each hole and incubated for 2 h in an incubator. The absorbance A value was measured at 490 nm using a Multiscan Spectrum microplate reader (Thermo, Waltham, MA, USA).

Wang P., Zhao Y., Fan R., Chen T, & Dong C. (2016). MicroRNA-21a-5p Functions on the Regulation of Melanogenesis by Targeting Sox5 in Mouse Skin Melanocytes. International Journal of Molecular Sciences, 17(7), 959.

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Quantifying EV-Associated Tissue Factor Activity

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The measurement of EV-associated TF activity was performed as previously described [19 (link)]. Briefly, after preparation, EVs were incubated with either an antibody for human TF (hTF1, 500μg/ml, 1μl; BD Biosciences, San Jose, CA, USA) or a control antibody (mouse IgG: 4 μg/ml; 1 μl; Sigma-Aldrich, St. Louis, 150 USA) and then 50 μl aliquots were added to duplicate wells of a 96-well plate. In the next step, 50 μl of HBSA containing 10 nM factor VIIa (FVIIa), 300 nM factor X (FX) and 10 mM CaCl₂ were added to each sample and the mixture incubated for 2 h at 37 °C. FXa generation was stopped by the addition of 25 μl of EDTA buffer and 25 μl of the chromogenic substrate Pefachrome FXa 8595 (4 mM; Pentapharm, Basel, Switzerland) were added and incubated at 37 °C for 15 min. Finally, absorbance at 405 nm was measured using a Multiscan Spectrum microplate reader (Thermo Scientific Inc., Bremen, Germany). EV-associated TF activity was calculated by reference to a standard curve that was generated using relipidated recombinant human TF. The TF-dependent FXa generation (pg/ml), which represents the EV-associated TF activity, was determined by subtracting the amount of FXa generated in the presence of hTF1 from the amount of FXa generated in the presence of the control antibody. Each measurement was performed in duplicates.

Hell L., Wisgrill L., Ay C., Spittler A., Schwameis M., Jilma B., Pabinger I., Altevogt P, & Thaler J. (2017). Procoagulant extracellular vesicles in amniotic fluid. Translational research : the journal of laboratory and clinical medicine, 184, 12-20.e1.

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Inflammatory Factors Quantification in HFLS

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After treatment for 48 h, HFLSs were harvested and centrifuged for 10 min at 400 × g, and inflammatory factor levels of IL-6 (cat. no. ab178013), IL-1β (cat. no. ab214025), TNF-α (cat. no. ab181421), MMP1 (cat. no. ab215083), and MMP3 (cat. no. ab269371) in the HFLS supernatant were quantified using ELISA kits (Abcam) following the manufacturer’s protocols. The OD value of the samples in each well was determined at 450 nm by a Multiscan Spectrum Microplate Reader (Thermo scientific), following the manufacturer’s protocol.

Ye Z., Wei L., Yin X., Li H., Qin G., Li S., Peng T., Liu B., Zhao S, & Zhuo Q. (2022). Long non-coding RNA cancer susceptibility candidate 2 regulates the function of human fibroblast-like synoviocytes via the microRNA-18a-5p/B-cell translocation gene 3 signaling axis in rheumatoid arthritis. Bioengineered, 13(2), 3240-3250.

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Microplate-based β-Galactosidase Assay

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β-Galactosidase was assayed using a Multiscan Spectrum Microplate Reader (Thermo Scientific). The absorbance at 420 nm was measured at 30°C immediately after the addition of ortho-nitrophenyl-β-galactoside substrate. Tester strains were incubated with shaking for 16 h at 28°C in 100 μl fresh CM medium and 100 μl of conditioned media produced by rhizoplane and riverbank isolates and harvested in T1. After the incubation, cells were centrifuged (4 °C; 1,800 × g) and re-suspended in 200 μl Z-buffer with 5.6% (vol/vol) β-mercaptoethanol before adding 10 μl toluene and incubating the cultures on ice for 30 min. The plate was then warmed to 30°C, 50 μl ortho-nitro-phenyl-β-galactoside substrate was added and the absorbance (420 nm) was immediately determined at 30°C.

Oslizlo A., Stefanic P., Vatovec S., Beigot Glaser S., Rupnik M, & Mandic-Mulec I. (2015). Exploring ComQXPA quorum-sensing diversity and biocontrol potential of Bacillus spp. isolates from tomato rhizoplane. Microbial Biotechnology, 8(3), 527-540.

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DPPH Radical Scavenging Assay Protocol

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The DPPH radical scavenging activity assay was performed following the procedure described by Sokolłetowska et al. [54 (link)] with some modifications. DPPH radical solution (200 μL of 0.2 mM) was added to a 50 μL aliquot of the 25-fold diluted extraction in a 96-well flat bottom microplate. After the mixture was mixed thoroughly and stored in the darkness for 20 min, the absorbance was measured using a multi-scan spectrum microplate reader (Thermo Scientific, Waltham, MA, USA) at 517 nm. A control containing 50 μL absolute ethanol was also included in each plate. The DPPH radical scavenging activity was calculated using Equation (2) with Trolox (0, 20, 40, 60, 80, 100, 120 and 140 μM), and results were expressed as μM Trolox equivalent (TE) per gram of fresh weight (μM TE/g fw).

DPPH radical scavenging rate = \frac{(A_{1} - A_{0}) - (A_{i} - A_{j})}{(A_{1} - A_{0})} \times 100 %

where A₁ and A_i represent the absorbance of solvent control and samples. A₀ and A_j represent the absorbance of the blank control and a blank sample.

Jia R., Tang C., Chen J., Zhang X, & Wang Z. (2022). Total Phenolics and Anthocyanins Contents and Antioxidant Activity in Four Different Aerial Parts of Leafy Sweet Potato (Ipomoea batatas L.). Molecules, 27(10), 3117.

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