To measure the fluorescence of each strain, colonies were inoculated into 180 μL of buffered minimal medium containing 4 g L−1 of glucose using BioscreenC (Growth Curves, Helsinki, Finland). After two rounds of subculture, the fluorescences of 100-μL samples were analyzed using the VICTOR3 1420 Multilabel Plate Reader (Perkin Elmer, MA, Waltham, USA). Specific fluorescence was calculated by dividing the fluorescence by the OD600. For inducible promoters, different concentrations of inducers were included at time zero (1 of mM IPTG, 100 μg L−1 of aTc, or 10 g L−1 of arabinose).
Victor3 1420 multilabel plate reader
Manufactured by PerkinElmer
Sourced in United States
The VICTOR3 1420 Multilabel Plate Reader is a versatile laboratory instrument designed for various detection methods. It can perform absorbance, fluorescence, and luminescence measurements in microplates.
Automatically generated - may contain errors
Lab products found in correlation
Cell counting kit 8 (cck8), by MultiSciences Biotech (1 mentions)
2 deoxyglucose, by Promega (1 mentions)
Glucose uptake glo assay, by Promega (1 mentions)
Palmitic acid, by Merck Group (1 mentions)
Insulin, by Merck Group (1 mentions)
Bactiter glo microbial cell viability assay, by Promega (1 mentions)
Quantichrom creatinine assay kit, by BioAssay Systems (1 mentions)
Chondroitin sulfate, by Merck Group (1 mentions)
Blyscan sulfated glycosaminoglycan sgag assay kit, by Biocolor (1 mentions)
7 protocols using victor3 1420 multilabel plate reader
1
Fluorescence Quantification of Microbial Strains
2
Cell Viability Assessment using CCK-8
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The Cell Counting Kit-8 (CCK-8) was purchased from MultiSciences Biotech Co. Ltd. (Hangzhou, China) and was used to assess the cultured DRG cell viability and cell proliferation. Cells were seeded in 96-well cell culture plates at a density of 2×104 cells/ml. The DRG cells were cultured with 10 μl CCK-8 in each well at 37°C for 1 h. Cell viability was measured using a VICTOR 31420 Multilabel plate reader (PerkinElmer, Waltham, MA, USA).
3
Palmitic Acid-Induced Insulin Resistance
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insulin resistance was induced using 0.5 mM palmitic acid (16:0, Sigma-Aldrich) for 48 h. Next, after IR induction, the appropriate concentration of CM fruit extract was added to the cells for an additional 48 h. Control cells were treated with DMSO. The day before the experiment, cells were starved in an FBS/FCS-free medium. On the day of the experiments, part of the cells was stimulated with 1 μM insulin (Sigma-Aldrich) for 15 min. Next, all cells were incubated with 1 mM of 2-deoxyglucose (as part of Glucose Uptake-Glo™ Assay, Promega Corporation) for an additional 15 min at RT. Then, the cells were further processed using a Glucose Uptake-Glo™ Assay according to the manufacturer’s protocol. The luminescence was measured using the Victor3, 1420 Multilabel Plate Reader (Perkin Elmer, Waltham, MA, USA). Glucose uptake in experimental and control cells was assessed based on three independent experiments.
4
Assaying IFN-γ production in tumor-specific CD8+ T cells
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For IFN-γ assays, 5e3 CD45.2+ CD3+ CD8 + T cells were sorted from each mouse brain and co-cultured with 100 μg/ml GL261-Luc2 tumor cell lysate and 25e3 dendritic cells isolated from CD45.1 mouse spleen (isolated with a pan-dendritic cell isolation kit) (Miltenyi Biotec) in a 96-well round bottomed plate with T cell media (RPMI 1640 + 10% FBS + 1% NEAA + 1% 2-Mercaptoethanol + 1% Penicillin/Streptomycin). Co-cultured cells were incubated at 37°C for 48 hours. Supernatant was collected for subsequent ELISA for IFN-γ and run on a plate reader (PerkinElmer Victor3 1420 Multilabel plate reader).
5
HA-H4R Expression Quantification
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Expression
of WT and mutant HA-H4R
on the surface of intact HEK293 cells was detected by ELISA using
1:800 anti-HA high-affinity clone 3F10 primary antibody (Sigma-Aldrich)
and 1:5000 horseradish peroxidase-conjugated goat anti-rat secondary
antibody (Thermo Fisher Scientific), as previously described.67 (link) Peroxidase activity was measured using 3,3′5,5′-tetramethylbenzidine
liquid substrate system (Abcam; Cambridge, UK) on a Victor3 1420 multilabel
plate reader (PerkinElmer) at 450 nm.
of WT and mutant HA-H4R
on the surface of intact HEK293 cells was detected by ELISA using
1:800 anti-HA high-affinity clone 3F10 primary antibody (Sigma-Aldrich)
and 1:5000 horseradish peroxidase-conjugated goat anti-rat secondary
antibody (Thermo Fisher Scientific), as previously described.67 (link) Peroxidase activity was measured using 3,3′5,5′-tetramethylbenzidine
liquid substrate system (Abcam; Cambridge, UK) on a Victor3 1420 multilabel
plate reader (PerkinElmer) at 450 nm.
6
Bactericidal Activity of Human Serum with C5 Inhibitors
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The bactericidal activity of human serum in the presence of C5 inhibitors was assessed using the BacTiter-Glo™microbial cell viability assay (Promega). Fifteen μl of NHS were incubated in PBS (final volume 80 μl) either alone or in the presence of increasing amounts of C5 inhibitor (C5:inhibitor molar ratios ranging from 1:1 to 1:1,500) for 2 h at 37°C under gentle agitation. In each tube was then added 40 μl of a culture of E. coli DH5α cells grown in LB medium at an 600OD (optical density at 600 nm) of 0.3. After 2 h of incubation at 37°C, each sample was aliquoted thrice in an opaque 96-well microplate (Nunc). An equivalent volume of BacTiter-GloTM solution was then added in each well and after 5 min. incubation at room temperature, the luminescence was read on a VICTOR 3 1420 Multilabel Plate Reader (PerkinElmer). The reference of 100% surviving bacteria was extrapolated from samples in which the DH5α cells were incubated with only PBS (no NHS). For each sample, the final read was obtained from averaging over the 3 replicate wells and the overall experiment was done in triplicate.
7
Quantification of Glycosaminoglycan Content
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All analyses were conducted in triplicate in a blinded manner. The quantification of glycosaminoglycan content involved assessing 100 μL aliquots of tissue homogenates and urine using the Blyscan–sulfated glycosaminoglycan (sGAG) assay kit (Biocolor Ltd., Carrickfergus, UK), following the manufacturer’s protocol. An aqueous solution of 10 mg/mL chondroitin sulfate (Sigma-Aldrich) was used as the reference standard. Absorbance was measured at 660 nm using a Victor3 1420 Multilabel Plate Reader (PerkinElmer, Waltham, MA, USA). Tissue GAG content was determined from the calibration curve (prepared with chondroitin sulfate amounts ranging from 0 to 5.0 μg in increments of 0.5–1.0 μg) and normalized for DNA concentration, measured using the Quanti-iT PicoGreen dsDNA Reagent (Thermo Fisher Scientific, Waltham, MA, USA) for tissue homogenates, or creatinine concentration, measured using the QuantiChrom Creatinine Assay Kit (BioAssay Systems, Hayward, CA, USA) for urine samples.
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