The cells were treated for 6 h with ligands with or without LPS. Consecutively, the total RNA was isolated using the RNeasy kit (QIAGEN, Hilden, Germany). Single-stranded cDNA was synthesized using the High Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific, Waltham, MA, USA). The qPCR was performed with a qPCR ABI 7500 Fast Real-Time PCR System (Applied Biosystems, Waltham, MA, USA) under usage of TaqMan™ Fast Advanced Master Mix (Thermo Fisher Scientific, Waltham, MA, USA). The following primers were used: (1) Hs00164383_m1, CYB1B1, FAM-MGB (2) Hs00174128_m1, TNFalpha, FAM-MGB (3) Hs00420895_gH, RPLPO, and FAM-MGB (all Thermo Fisher, USA). Fold changes were calculated by the ΔΔct-method relative to the unstimulated control with RPLP0 as the reference [29 (link)].
Rplpo
Manufactured by Thermo Fisher Scientific
Sourced in United States
The RPLPO is a laboratory instrument designed to detect and quantify the expression of the RPLP0 gene, which is commonly used as a reference gene in gene expression analysis. The RPLPO measures the levels of this gene transcript using a standardized protocol, providing researchers with accurate and reliable data to support their scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (4 mentions)
Hs00420895 gh, by Thermo Fisher Scientific (1 mentions)
Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions)
Fam mgb, by Thermo Fisher Scientific (1 mentions)
Hs00174128 m1, by Thermo Fisher Scientific (1 mentions)
Qpcr abi 7500 fast real time pcr system, by Thermo Fisher Scientific (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Rneasy kit, by Qiagen (1 mentions)
Gapdh, by Thermo Fisher Scientific (1 mentions)
Steponeplus, by Thermo Fisher Scientific (1 mentions)
Direct zol rna miniprep plus kit, by Zymo Research (1 mentions)
2 protocols using rplpo
1
Gene Expression Analysis of Immune Responses
2
Quantifying Gene Expression under Hypoxia
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The RT-qPCR analysis was performed on StepOne plus (Applied Biosystems). Total RNA was extracted from hypoxic and normoxic cells using a Direct-Zol RNA MiniPrep plus Kit (Zymo Research), cDNA was synthesized using a high capacity cDNA reverse transcription kit (Thermo Fisher Scientific), and a
TaqMan probe (PCR Master Mix -Life Technologies)
was used for the PCR reaction. Amplification of two housekeeping genes, GAPDH (Thermo Fisher Scientific)
and RPLPO (Thermo Fisher Scientific), were used as an internal standard. The comparative Ct method was applied to determine the fold-change in expression levels relative to the control samples, according to the formula ΔΔCt (2 - ΔΔCt ) [16] .
TaqMan probe (PCR Master Mix -Life Technologies)
was used for the PCR reaction. Amplification of two housekeeping genes, GAPDH (Thermo Fisher Scientific)
and RPLPO (Thermo Fisher Scientific), were used as an internal standard. The comparative Ct method was applied to determine the fold-change in expression levels relative to the control samples, according to the formula ΔΔCt (2 - ΔΔCt ) [16] .
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