NF-κB or IRF activation was analyzed by secreted embryonic alkaline phosphatase (SEAP) (QUANTI-Blue™, InvivoGen, rep-qbs) or Lucia luciferase (QUANTI-Luc™, InvivoGen, rep-qlc1) reporter assay in cell supernatants after stimulation with 10 ng/mL LPS for 24 hr, according to the manufacturer's instructions. QUANTI-Luc™ was analyzed immediately by luminescence and QUANTI-Blue™ after 45 min of incubation by absorbance at 635 nm on a Synergy HTX S1LFA plate reader (BioTek).
Synergy htx s1lfa plate reader
Manufactured by Agilent Technologies
The Synergy HTX S1LFA plate reader is a compact and versatile microplate reader from Agilent Technologies. It is designed to perform label-free, real-time detection of molecular interactions using the Symphony platform's advanced optics and detection capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Quanti blue, by InvivoGen (1 mentions)
Quanti blue, by Agilent Technologies (1 mentions)
Quanti luc, by InvivoGen (1 mentions)
Quick rna miniprep kit, by Zymo Research (1 mentions)
Iscript cdna kit, by Bio-Rad (1 mentions)
Primepcr probe assay, by Bio-Rad (1 mentions)
Ssoadvanced universal probes supermix, by Bio-Rad (1 mentions)
Cfx96 real time pcr system, by Bio-Rad (1 mentions)
Quantifluor rna system, by Promega (1 mentions)
2 protocols using synergy htx s1lfa plate reader
1
NF-κB/IRF Activation Analysis
2
Quantitative Real-Time PCR Analysis
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Cells were lysed and RNA was isolated by the Quick-RNA Miniprep Kit (Zymo Research, R1054). RNA concentration was measured by QuantiFluor® RNA System (Promega) on a Synergy HTX S1LFA plate reader (BioTek). cDNA was synthesized by the iScript cDNA kit (BioRad, 1708890) and run with the PrimePCR™ Probe Assays (BioRad) for human TNF (qHsaCEP0040184), CXCL10 (qHsaCEP0053880), IFNB1 (qHsaCEP0054112), HPRT1 (qHsaCIP0030549), and B2M (qHsaCIP0029872) and the SsoAdvanced™ Universal Probes Supermix (BioRad, 1725280) on a CFX96 Real-Time PCR System (BioRad). The mean of HPRT and B2M Cts was used for normalization. Relative mRNA levels were transformed into a linear form by the 2 (−ΔΔCt) method [41 (link)]. Fold changes of RNA expression relative to unstimulated controls were used for statistical analysis.
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