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Ifna4 is a laboratory equipment product from Thermo Fisher Scientific. It is a specialized device used for conducting scientific research and analysis. The core function of Ifna4 is to provide a controlled and precise environment for conducting experiments and testing procedures. However, a detailed description of its intended use or specific applications cannot be provided while maintaining an unbiased and factual approach.

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3 protocols using ifna4

1

Quantifying Type I Interferons in Lymph Nodes

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Total RNA was isolated from frozen LNs with ReliaPrep RNA Miniprep system (Promega), following the manufacturer’s instructions. 1 μg of total RNA was reverse transcribed prior to qPCR analyses for Ifna2 (Mm00833961_S1, Thermo Fisher), Ifna4 (Mm00833969_S1, Thermo Fisher), Ifna5 (Mm00833976_S1, Thermo Fisher), Ifna6 (Mm01703458_S1, Thermo Fisher), Ifna7 (Mm02525960_S1, Thermo Fisher), Ifna9 (Mm00833983_S1, Thermo Fisher), Ifna12 (Mm00616656_S1, Thermo Fisher), Ifna13 (Mm01731013_S1, Thermo Fisher), Ifna14 (Mm01703465_S1, Thermo Fisher), Ifnb (Mm00439552_S1, Thermo Fisher), Isg15 (Mm 01705338_S1,hermo Fisher), 2’5’Oas (Mm00460961_m1, Thermo Fisher) and IL-6 (Mm00446190_M1, Thermo Fisher) in a QuantStudio 5 Real-Time PCR System (Thermo Fisher Scientific). All experiments were done in duplicate and normalized to the housekeeping gene GAPDH (Mm99999915_g1, Thermo Fisher).
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2

Quantitative PCR analysis of human liver

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Whole liver RNA was isolated using RNeasy mini kit (Qiagen, Hilden, Germany) including DNAse treatment according to manufacturer’s protocol starting with homogenization of liver tissue in RLT buffer. cDNA was generated by using an iScript cDNA synthesis kit (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s protocol. Human specific gene expression was measured using Taqman primer/probe quantitative PCR, in TaqMan® Gene Expression Master Mix (Thermo Fisher Scientific, Waltham, MA, USA). Primer/probe combinations were purchased from Thermo Fisher Scientific; CXCL10 (Hs01124251_g1), CXCL9 (Hs00171065_m1), DDX58 (Hs01061436_m1), GAPDH (Hs00266705_g1), IFIT1 (Hs01911452_s1), ISG15 (Hs01921425_s1), IFNA1 (Hs00855471_g1), IFNA4 (Hs01681284_sh), IFNB1 (Hs01077958_s1), MX1 (Hs00895608_m1), OAS1 (Hs00973637_m1), RSAD2 (Hs00369813_m1), STAT1 (Hs01013996_m1), TLR3 (Hs01551078_m1). Expression of target genes was normalized to the expression of GAPDH using the formula 2−ΔCt, ΔCt = Cttarget−CtGADPH. cDNA from non-chimeric mouse livers was used as control to test cross-reactivity of housekeeping and target genes. Due to the difference in hepatocyte donor baseline expression levels of examined genes (Suppl. Fig. 1), fold changes of transcripts were calculated to those of non-infected humanized livers from mice transplanted with the identical hepatocyte donor.
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3

Quantifying Type I Interferons in Lymph Nodes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was isolated from frozen LNs with ReliaPrep RNA Miniprep system (Promega), following the manufacturer’s instructions. 1 μg of total RNA was reverse transcribed prior to qPCR analyses for Ifna2 (Mm00833961_S1, Thermo Fisher), Ifna4 (Mm00833969_S1, Thermo Fisher), Ifna5 (Mm00833976_S1, Thermo Fisher), Ifna6 (Mm01703458_S1, Thermo Fisher), Ifna7 (Mm02525960_S1, Thermo Fisher), Ifna9 (Mm00833983_S1, Thermo Fisher), Ifna12 (Mm00616656_S1, Thermo Fisher), Ifna13 (Mm01731013_S1, Thermo Fisher), Ifna14 (Mm01703465_S1, Thermo Fisher), Ifnb (Mm00439552_S1, Thermo Fisher), Isg15 (Mm 01705338_S1,hermo Fisher), 2’5’Oas (Mm00460961_m1, Thermo Fisher) and IL-6 (Mm00446190_M1, Thermo Fisher) in a QuantStudio 5 Real-Time PCR System (Thermo Fisher Scientific). All experiments were done in duplicate and normalized to the housekeeping gene GAPDH (Mm99999915_g1, Thermo Fisher).
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