Interleukin (IL)-6 expression and secretion was quantified using the BD OptEIA Human IL6 ELISA kit II (BD Biosciences, CA, USA) according to the manufacturer’s protocol. Briefly, confluent HBMEC were treated with PIC (50 µl/ml), HIV-1ADA (MOI: 0.001) and/or pharmacological inhibitors (Table 3 ) for 48 hours. Controls consisted of untreated cells and cells treated with the TLR3/dsRNA Complex Inhibitor (TLR3.CI) (30 nM). Following cell treatment, culture supernatants were collected and further centrifuged for 5 min at 2350 g to remove any cellular debris, and 100 µl of each sample used for IL6 ELISA according to manufacturer’s instructions. Absorbance readings were obtained using SpectraMax M5 ELISA plate reader (Molecular Devices, Sunnyvale, CA, USA). For each experiment, IL6 standards provided with the kit were used to generate a standard curve and determine IL6 concentrations in each sample. Each experimental condition was tested in duplicate or triplicate.
Spectramax m5 elisa
Manufactured by Molecular Devices
Sourced in United States
The SpectraMax M5 ELISA is a multi-mode microplate reader designed for a variety of detection methods, including absorbance, fluorescence, and luminescence. It is capable of performing enzyme-linked immunosorbent assays (ELISAs) and other microplate-based assays.
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Lab products found in correlation
3 protocols using spectramax m5 elisa
1
Quantifying IL-6 Expression and Secretion in HBMEC
2
Quantifying Breast Cancer Cell Proliferation
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The proliferation of each breast cancer cell line was monitored using an MTT Cell Proliferation/Viability Assay kit (R&D Systems, Inc., Minneapolis, MN, USA), according to the manufacturer’s protocol. Briefly, following transfection with si-hsa_circ_0011946 or si-NC, 15 µL of MTT was added to each well (5×104) and incubated for 4 h at 37°C. Then, the stop solution was added and the solution was kept overnight at room temperature. The optical density value was measured at 492 nm using a SpectraMax M5 ELISA plate reader (Molecular Devices, LLC, Sunnyvale, CA, USA).
3
Quantification of Fluorescent AGEs
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Liver, fat, and serum fluorescent AGEs were quantified using the Nakayama method.8 (link),10 (link),43 (link) Sample fluorescence was measured against blank (0.1N NaOH) at an emission wavelength of 440 nm and an excitation wavelength of 370 nm using a SpectraMax M5 ELISA microplate reader (Molecular Devices, USA). One fluorescence unit was equivalent to the fluorescence intensity of 1 mg/mL of native bovine serum albumin (BSA) and expressed as arbitrary units (AU).
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