Total RNA was extracted from mTEC. Real-time PCR was performed with a machine (Option 2, Bio-Rad, Hercules, CA, USA) by using the IQ SYBR Green Supermix reagent (Bio-Rad). The primers used in this study were as follows: mouse IL-1β, forward 5′-TGCCACCTTTTGACAGTGATG-3′ and reverse 5′-AAGGTCCACGGGAAAGACAC-3′; TNF-α, forward 5′-CATCTTCTCAAAATTCGAGTGACAA-3′ and reverse 5′-TGGGAGTAGACAAGGTACAACCC-3′ [11 (link),12 (link)]. The housekeeping gene β-actin was used as an internal control. The ratio of the specific mRNA: β-actin mRNA was calculated using the 2−ΔCt method.
Option 2
Manufactured by Bio-Rad
Sourced in United States
The Option 2 lab equipment from Bio-Rad is a versatile instrument designed for a range of laboratory applications. It features a compact and durable construction, allowing for efficient use of space in the laboratory setting. The core function of this product is to perform essential tasks required for various scientific experiments and analyses.
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Lab products found in correlation
3 protocols using option 2
1
Quantitative Analysis of Cytokine Expression in Mouse TECs
2
Real-Time PCR for Gene Expression Analysis
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Total RNA was extracted from the renal cortical tissues and cultured HK-2 cells and real-time PCR was performed with a real-time PCR machine (Option 2, Bio-Rad, Hercules, CA, USA) by using IQ SYBR green supermix reagent (Bio-Rad)47 (link)48 . The primers for mouse mRNA MCP-1, IL-1β, TNFa, collagen I, collagen IV, CD32 and human mRNA CTGF have been described previously9 (link)10 48 49 (link), The housekeeping genes β-actin was used as internal controls. The ratio of specific mRNA: β-actin mRNA was calculated using the 2−ΔΔCt method and is expressed as the mean ± S.E.M.
3
Quantitative Real-Time PCR Analysis of Renal Gene Expression
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Total RNA was extracted from the renal cortical tissues and cultured renal cell lines. Real-time PCR was carried out with machine (Option 2, Bio-Rad, Hercules, CA, USA) by using IQ SYBR Green Supermix reagent (Bio-Rad) 16 (link), 17 . The Sequence of RNA Primers used such as Smad3, Smad7, collagen-I, α-SMA, MCP-1, NF-kB, ERBB4-IR, LRNA9884, and GAPDH were described previously 5 (link), 7 (link), 12 (link), 18 (link). The house keeping genes β-actin was used as internal controls. The ratio of specific mRNA to β-actin mRNA was calculated using the 2-ΔCt method and is expressed as the mean ± S.E.M.
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