Anti igg hrp alexseries

Protein samples were prepared by mixing one part of sample with one part of Bio-Rad Sample Buffer and then boiled at 100 °C for 5 min. Proteins were separated in 8–12% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane at 200 mA for 2 h in ice treatment. The blot was blocked with 5% fat-free dry milk suspended in 1× TBST for 15 min at RT. Resulting blot was incubated with antibodies (1:500–1:1000), including connexin43/β-catenin/active β-catenin/lef-1 for overnight at 4 °C, followed by incubation with 1:2000 anti-IgG-HRP (AlexSeries, Abcam) for 2 h at RT. Signals from the blots were obtained using a Western Blotting Luminol Reagent Kit.

Protein samples were prepared by mixing one part of sample with one part of Bio-Rad Laemmli Sample Buffer and then boiled at 100 °C for 5 min. Proteins were separated in 8%–12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (according to the molecular weights of the LOX profile) and transferred to a polyvinylidene difluoride membrane at 200 mA for 1 h at RT. The blot was blocked with 5% fat-free dry milk suspended in 1× TBST for 2 h at RT. The resulting blot was incubated with antibodies (1:500–1 000), including β-catenin/phosphorylated β-catenin, GAPDH, VEGFA, Lef-1, and cyclin D (Abcam), for 1–3 h at RT, followed by incubation with 1:5 000 anti-IgG-HRP (Alex Series, Abcam) for 2 h at RT. Signals from the blots were obtained using a Western Blotting Luminol Reagent Kit. Proteins were visualized via chemiluminescence with hydrogen peroxide using Kodak X-AR and luminol as substrates.