20’000 LL171 cells/well were grown in a 96 well flat bottom luciferase reading plate over night in 10% RPMI medium supplemented 1% L-glutamine (all Gibco). On the next day, 10-fold pre-diluted test sera and respective IFN-β standard (Biolegend) were added in RPMI medium and incubated for 24 h at 37 °C. The cells were then lysed in 1x lysis buffer (Promega) for 10 min, followed by the addition of luciferase substrate (Promega) via injector system. The luciferase activity was measured on a Tecan Infinite 200 Pro plate reader.
Ifn β standard
Manufactured by BioLegend
The IFN-β standard is a recombinant protein used as a reference standard for the quantification of interferon-beta (IFN-β) in biological samples. It serves as a calibrator for IFN-β measurement assays.
Automatically generated - may contain errors
Lab products found in correlation
3 3 5 5 tetramethylbenzidine (tmb), by Thermo Fisher Scientific (2 mentions)
Avidin hrp, by Thermo Fisher Scientific (2 mentions)
Purified anti mouse ifn β antibody, by BioLegend (2 mentions)
Biotin anti mouse ifn β antibody, by BioLegend (2 mentions)
Lysis buffer, by Promega (1 mentions)
Luciferase substrate, by Promega (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Infinite 200 pro, by Tecan (1 mentions)
Ifn gamma elisa kit, by Thermo Fisher Scientific (1 mentions)
Mouse elisa kit, by Thermo Fisher Scientific (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
3 protocols using ifn β standard
1
Luciferase Assay for IFN-β Activity
2
IFN-β and IFN-γ Measurement in M. avium Infection
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IFN-β production in mouse serum or BMMs culture supernatants at 24 or 72 hr post M.avium infection at 37°C and 5% CO2 was measured. ELISA was performed according to the manufacturer’s instruction (ebioscience). Capture antibody (Purified anti-mouse IFN-β Antibody, Cat. 519202, Biolegend), detection antibody (Biotin anti-mouse IFN-β Antibody, Cat. 508105, Biolegend), IFN-β standard (Cat. 581309, Biolegend), Avidin-HRP (Cat.18-4100-94, ebioscience), and TMB (Cat.00-4201-56, ebioscience). IFN-γ in culture supernatant and lung homogenate was measured using IFN gamma ELISA kit (Cat. 88-7314-22; eBioscience). For in vivo IFN-γ measurement, mouse lung homogenate was prepared as described previously [35 (link)].
3
Cytokine Quantification in Macrophage Infection
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Macrophage culture supernatants were used to measure IFN-β and TNFα level at 24 h after M.tb infection. Culture supernatants were centrifuged at 1,000 xg for 10 min at 4°C and subsequently passed through 0.2-micron filter two times. The ELISA was performed according to the manufacturer’s protocol (eBioscience) using Avidin-HRP (cat. 18-4100-94; eBioscience) and TMB (cat. 00-4201-56; eBioscience). For TNFα, capture antibody (cat. 14-7341-85; eBioscience), detection antibody (cat. 13-7341-85; ebioscience), and TNFα standard (cat. 39-8321-60; eBioscience) were used. For IFN-β, capture antibody (purified anti-mouse IFN-β antibody, cat. 519202; BioLegend), detection antibody (biotin anti-mouse IFN-β antibody, cat. 508105; BioLegend), and IFN-β standard (cat. 581309; BioLegend) were used. Mouse IFN-γ and IL-1β were measured using IFN gamma (cat. 88-7314-22; eBioscience) and IL-1 β (cat. 88-7013-22; eBioscience) mouse ELISA kit, respectively.
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