RNA was isolated from cells using the RNeasy plus Kit (Qiagen, Hilden, Germany) following the manufacturers’ instructions. 100 ng RNA were used for the real-time PCR reaction using the QuantiTect SYBR Green RT-PCR Kit from Qiagen following the manufacturer’s instructions. Analysis was performed using the CFX96 Touch Real-Time PCR Detection System (Bio-RAD, Hercules, CA, USA). The following QuantiTect primers were used: Hs_GAPDH_2_SG , Hs_PLCE1_1_SG, Hs_ARHGEF2_1_SG, Hs_PRKD2_1_SG, Hs_PRKD3_1_SG, and Hs_ACTB_2_SG (all obtained from Qiagen).
Hs actb 2 sg
Manufactured by Qiagen
Sourced in United Kingdom
Hs_ACTB_2_SG is a real-time PCR assay designed to detect the human beta-actin (ACTB) gene. It is a core reference gene that is commonly used for gene expression normalization in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Quantitect sybr green rt pcr kit, by Qiagen (1 mentions)
Rneasy plus kit, by Qiagen (1 mentions)
Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions)
Hs gapdh 2 sg, by Qiagen (1 mentions)
Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions)
Tissuelyser 2, by Qiagen (1 mentions)
Nanodrop 2000, by Thermo Fisher Scientific (1 mentions)
Rneasy kit, by Qiagen (1 mentions)
Omniscript rt kit, by Qiagen (1 mentions)
Tripure, by Roche (1 mentions)
Viia 7 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Direct zol rna miniprep kit, by Zymo Research (1 mentions)
Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions)
Quantitect primer, by Qiagen (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Fast sybr green master mix, by Thermo Fisher Scientific (1 mentions)
3 protocols using hs actb 2 sg
1
Comprehensive RNA Extraction and qPCR Analysis
2
RNA Isolation and qRT-PCR Analysis
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RNA isolation of human cells was performed in RLT lysis buffer working solution containing 1% β-mercaptoethanol according to manufacturer's protocols. Murine aortas were lysed in 1 ml Tripure (Roche, Mannheim, Germany) by a tissuelyser II (Qiagen, Hilden, Germany) for 5 min at 29 s−1 for mRNA isolation. mRNA purification of cells and aortas was done with an RNeasy Kit (Qiagen, Hilden, Germany) and content of mRNA was measured with a NanoDrop2000 (Thermo Scientific, Schwerte, Germany). All samples were reversely transcribed into 1 μg/μl of cDNA using an Omniscript RT Kit (Qiagen, Venlo, Netherlands) and mRNA levels were determined by StepOne Plus real-time PCR system (Applied Biosystems, Carlsbad, CA, USA). Primers from Qiagen were used: Hs_F2RL1_1_SG (QT02589377), Hs_ACTB_2_SG (QT01680476) Hs_PTGS2_SG (QF00461055), mM_F2RL1 (QT02255330), and Mm_Rn18S (QT02448075).
3
Quantitative Analysis of Matrix Metalloproteinases
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RNA was extracted in TRIzol (Invitrogen), purified (Direct-Zol RNA miniprep kit; Zymo Research, Cambridge, UK) and reverse transcribed (High capacity cDNA reverse transcription kit; Applied Biosystems, California, USA). Quantitative PCR was performed using Fast SYBR Green Master Mix or TaqMan Fast Advanced Master Mix in a Viia7 Real-Time PCR system (Applied Biosystems, Warrington, UK). Human primers were either pre-validated Quantitect primers against MMP1 (Hs_MMP1_1_SG) , MMP3 (Hs_MMP3_1_SG), MMP9 (Hs_MMP9_1_SG) , MMP13 (Hs_MMP13_1_SG) or ACTB (Hs_ACTB_2_SG) (Qiagen, Manchester, UK), Primerdesign primers against MMP8 (JN215643) or ADAMTS9 (JN171532) (Primerdesign, Chandlers Ford, UK) or designed in-house (Supplementary Table 1). A gene was not considered to be expressed when the average cycle threshold (Ct) value per condition was higher than 34. The expression of individual mRNAs was calculated relative to expression of -actin (ACTB) using the (2-ΔCt) method.
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