It was determined using quantitative real-time PCR after total RNA was isolated according to the manufacturer’s instructions using the Qiagen tissue extraction kit (Qiagen, USA). Using a high-capacity cDNA reverse transcription kit (Fermentas, USA), total RNA was converted to cDNA. Then, using Applied Biosystems with Step One TM software version 3.1 (USA), amplification and analysis of real-time qPCR product were conducted. The primer sequence of the gene under study include:
Step one software version 3
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Step One TM software version 3.1 is a data analysis tool designed for use with Thermo Fisher Scientific's real-time PCR instruments. The software provides a user-friendly interface for analyzing and interpreting data generated from real-time PCR experiments.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using step one software version 3
1
Quantitative Real-Time PCR Protocol
2
Multimodal Evaluation of Renal Inflammation
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Colorimetric method was used determine SOD activity ( Sun et al. 1988 ).
Quantitative estimation of tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL6) concentration in renal tissue:
It was assessed using USCN life Science Inc. ELISA kits. The competitive inhibition enzyme immunoassay technique is used in this assay.
Estimation Bcl-2 associated x protein (Bax) and nuclear factor kappa B (NFκB) in renal tissue:
It was determined using quantitative real-time PCR after total RNA was isolated according to the manufacturer's instructions using the Qiagen tissue extraction kit (Qiagen, USA). Using a high-capacity cDNA reverse transcription kit (Fermentas, USA), total RNA was converted to cDNA. Using Applied Biosystems with Step One TM software version 3.1 (USA), ampli cation and analysis of the real-time qPCR product were performed. The primer sequence of the studied genes; BAX: Forward primer:5'-CCCTGTGCACTAAAGTGCCC-3. Reverse primer: 5'-CTTCTTCACGATGGTGAGCG-3 NFκB: Forward primer:5'-CATTGAGGTGTATTTCACGG -3
Reverse primer: 5'-GGCAAGTGGCCATTGTGTTC -3
Quantitative estimation of tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL6) concentration in renal tissue:
It was assessed using USCN life Science Inc. ELISA kits. The competitive inhibition enzyme immunoassay technique is used in this assay.
Estimation Bcl-2 associated x protein (Bax) and nuclear factor kappa B (NFκB) in renal tissue:
It was determined using quantitative real-time PCR after total RNA was isolated according to the manufacturer's instructions using the Qiagen tissue extraction kit (Qiagen, USA). Using a high-capacity cDNA reverse transcription kit (Fermentas, USA), total RNA was converted to cDNA. Using Applied Biosystems with Step One TM software version 3.1 (USA), ampli cation and analysis of the real-time qPCR product were performed. The primer sequence of the studied genes; BAX: Forward primer:5'-CCCTGTGCACTAAAGTGCCC-3. Reverse primer: 5'-CTTCTTCACGATGGTGAGCG-3 NFκB: Forward primer:5'-CATTGAGGTGTATTTCACGG -3
Reverse primer: 5'-GGCAAGTGGCCATTGTGTTC -3
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