About Me:I was born in Southwestern Illinois, moved to Madison when I was six, and have lived here ever since. Both of my parents are teachers (though my dad is doing something else right now, but he's still a teacher, just not in a school) and that's something that I know how ...
RNase-free DNase was from Promega Corporation (Madison, WI). All the other reagents were from Sigma or as indicated in the specified methods. Animals and treatments. The ICR mice (8~10 week-old; male mice: 28~30 g; female mice: 24~26 g) were purchased from Beijing Vital ...
RAW 264.7 cells in 24-well plates were transfected in triplicate with the NF-κB promoter-driven luciferase reporter plasmid pNF-κB-Luc (200 ng/well) and pRL-TK-Luc (5 ng/well) by using FuGENE HD transfection reagent (Promega, Madison, WI, USA). Twenty four hours later, cells were sti...
(TBST) for 2 h, then were incubated overnight with the corresponding antibodies at 4 °C, followed by incubation with horseradish peroxidase-conjugated goat anti-mouse IgG (W402B, Promega, Madison, WI, USA) or goat anti-rabbit IgG (W401B, Promega), or rabbit anti-Goat IgG (Zymax...
(TBST) for 2 h, then were incubated overnight with the corresponding antibodies at 4 °C, followed by incubation with horseradish peroxidase-conjugated goat anti-mouse IgG (W402B, Promega, Madison, WI, USA) or goat anti-rabbit IgG (W401B, Promega), or rabbit anti-Goat IgG (Zymax...
Advanced glycated end products (AGEs) are cytotoxic compounds that are mainly increased in diabetes mellitus (DM), kidney failure, inflammation, and in response to the ingestion of AGE-rich diets. AGEs can also impair glycemic homeostasis by decreasing t
Subsequently, 20 μL MTS reagent (Cell Titer 96 Aqueous One Solution Cell Proliferation Assay, Promega, Madison, WI, USA) was pipetted into each well and mixed thoroughly. After 20 min incubation at 37 °C and 5% CO2, 80 μL supernatant of each well was transferred into the wells of a...
Exactly 1 μg of RNA sample was used to synthesize cDNA by reverse transcriptase reaction using M-MLV reverse transcriptase kit (Promega, Madison, WI, USA). cDNA synthesis followed a two-step reaction condition of 70 °C for 5 min and 42 °C for 1 h. Expression of mRNA was quantified ...
[52]. The nuclear proteins from the transfected cells were then dissolved in 20 mM Hepes (pH 7.9) containing 0.4 M NaCl, 1 mM EDTA, 1 mM EGTA, 1 mM DTT, 1 mM PMSF and a cocktail of protease inhibitors (Promega Corp, Madison, WI, USA). The protein concentration of each extract ...
The nitrite accumulated in the culture medium was measured as an indicator of NO production using the Griess reagent kit (Promega, Madison, WI, USA). Cell culture media were centrifuged at 4 °C for 5 min and the supernatant was subjected to measurement of nitrite. ...