Hrp conjugated affinipure goat anti rabbit igg for dsg2 and gapdh

Jejunum tissues were lysed with radioimmunoprecipitation assay (RIPA) buffer (Beyotime, China). Proteins were electrophoretically separated on SDS-PAGE gels and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, USA). After blocking with 5% nonfat milk, the membranes were incubated with anti-KLF5 (1:500; Origene, USA) or anti-DSG2 (1:1,000; Thermo Fisher, USA) primary antibodies overnight at 4°C. After washing, the specific blots were incubated with the species-appropriate secondary antibodies HRP-conjugated AffiniPure goat anti-mouse IgG for KLF5 (1:5,000; Boster, China) and HRP-conjugated AffiniPure goat anti-rabbit IgG for DSG2 and GAPDH (1:5,000; Boster, China) for 1 h at room temperature. Finally, the protein bands were viewed with an Imaging System ChemiDoc MP (Bio-Rad, USA) and then analyzed with ImageJ software. In the quantitative analysis of Western blots, all of the bands detected were within the linear range of detection.

Jejunum tissues were lysed with radioimmunoprecipitation assay (RIPA) buffer (Beyotime, China). Proteins were electrophoretically separated on SDS-PAGE gels and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, USA). After blocking with 5% nonfat milk, the membranes were incubated with anti-KLF5 (1:500; Origene, USA) or anti-DSG2 (1:1,000; Thermo Fisher, USA) primary antibodies overnight at 4°C. After washing, the specific blots were incubated with the species-appropriate secondary antibodies HRP-conjugated AffiniPure goat anti-mouse IgG for KLF5 (1:5,000; Boster, China) and HRP-conjugated AffiniPure goat anti-rabbit IgG for DSG2 and GAPDH (1:5,000; Boster, China) for 1 h at room temperature. Finally, the protein bands were viewed with an Imaging System ChemiDoc MP (Bio-Rad, USA) and then analyzed with ImageJ software. In the quantitative analysis of Western blots, all of the bands detected were within the linear range of detection.