Twelve days after immunization with IRBP 1–20 peptide, mice (n = 5/group) were sacrificed and their spleen collected and dissociated. Splenic cells were proceeded for qRT-PCR analysis of CD3 and Foxp3 gene expression. β-actin was used as a non-modulated reference gene. The mRNA extraction and isolation were carried out using the automated MagNA Pure LC Instrument system and the MagNAPure LC mRNA Isolation Kit II, following manufacturer’s instructions (Roche Applied Science, Vilvoorde, Belgium). A one step real-time quantitative PCR technique using the RNA Master Hybridization Probes Kit (Roche Applied Science) was used to quantify mRNA expression using specific primes and fluorescent probes for mouse CD3, Foxp3 and β-actin (Applied Biosystems). Data are presented as relative expression of Foxp3 versus CD3.
Magnapure lc mrna isolation kit 2
Manufactured by Roche
Sourced in Belgium
The MagNAPure LC mRNA Isolation Kit II is a laboratory equipment product designed for the isolation of messenger RNA (mRNA) from various sample types. It utilizes magnetic bead-based technology to capture and purify mRNA molecules from cellular samples.
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2 protocols using magnapure lc mrna isolation kit 2
1
Splenic T-cell Gene Expression after IRBP Immunization
2
SOCS1 Regulation by Inflammatory Cytokines
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ARPE-19 cells, CTRL clone or SOCS1 clone were seeded at 5 × 10 5 cells/well in 6-well plates. One day later, IFNγ (150 ng/ml), TNFα (30 ng/ml) or IL-17 (100 ng/ml) were added to the cultures for 24 hours. Cells were collected for qRT-PCR analysis of SOCS1 gene expression. β-Actin was used as a non-modulated reference gene. The mRNA extraction and isolation were carried out using the automated MagNA Pure LC Instrument system (LightCycler® RNA Master) and the MagNAPure LC mRNA Isolation Kit II, following manufacturer's instructions (Roche Applied Science, Vilvoorde, Belgium). A one step real-time quantitative RT-PCR technique using the RNA Master Hybridization Probes Kit (Roche Applied Science) was used to quantify mRNA expression using specific primes and fluorescent probes for human SOCS1 (Primer and probe SOCS1 human, Applied Biosystems SOCS1 TaqMan) and β-actin (Primer and probe B-actine human, Applied Biosystems, TaqMan Gene Expression Assays).Data are presented as normalized expression of SOCS1 versus β-actin (2 -(CtSOCS1-Ctβactin) ).
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