Rabbit anti goat igg hrp antibodies

The tissues were immediately fixed for 24 h in freshly prepared 4% paraformaldehyde in PBS, processed, and paraffin-embedded. Three-micrometer sections were washed in xylene and rehydrated in gradient ethanol aliquots. Sections were stained with hematoxylin-eosin (H&E). An optical microscope was used to observe and photograph the tissues. Tissue images were evaluated using an optical microscope (Olympus Corporation, Tokyo, Japan). The villus height of the jejunum and ileum was measured using H&E images. Paraformaldehyde-fixed, paraffin-embedded sections were heated with 10 mmol/L citric buffer at 121 °C for ten minutes, incubated with 3% H₂O₂ /PBS and 0.5%Triton/PBS, blocked with Protein Block (Agilent Technologies, Santa Clara, CA, USA), stained with Goat anti–IL-13 polyclonal antibodies (Santa Cruz Biotechnology, Dallas, TX, USA) and Rabbit anti-goat IgG-HRP antibodies (Santa Cruz Biotechnology), followed by the use of the ImmPACT DAB Substrate Kit (Vector Laboratories, Inc., Newark, CA, USA). The percentage of IL-13-positive areas was calculated using the villus area, with the muscle layer excluded as the denominator. The quantification was performed using Image J (NIH, Bethesda, MD, USA). IL-13 and CD4 double staining were performed using anti-IL-13 IgG-PE and anti-CD4 IgG-FITC antibodies (Thermo Fisher Scientific Inc., Waltham, MA, USA).

The tissues were immediately fixed for 24 h in freshly prepared 4% paraformaldehyde in PBS, processed, and paraffin-embedded. Three-micrometer sections were washed in xylene and rehydrated in gradient ethanol aliquots. Sections were stained with hematoxylin-eosin (H&E). An optical microscope was used to observe and photograph the tissues. Tissue images were evaluated using an optical microscope (Olympus Corporation, Tokyo, Japan). The villus height of the jejunum and ileum was measured using H&E images. Paraformaldehyde-fixed, paraffin-embedded sections were heated with 10 mmol/L citric buffer at 121 °C for ten minutes, incubated with 3% H₂O₂ /PBS and 0.5%Triton/PBS, blocked with Protein Block (Agilent Technologies, Santa Clara, CA, USA), stained with Goat anti–IL-13 polyclonal antibodies (Santa Cruz Biotechnology, Dallas, TX, USA) and Rabbit anti-goat IgG-HRP antibodies (Santa Cruz Biotechnology), followed by the use of the ImmPACT DAB Substrate Kit (Vector Laboratories, Inc., Newark, CA, USA). The percentage of IL-13-positive areas was calculated using the villus area, with the muscle layer excluded as the denominator. The quantification was performed using Image J (NIH, Bethesda, MD, USA). IL-13 and CD4 double staining were performed using anti-IL-13 IgG-PE and anti-CD4 IgG-FITC antibodies (Thermo Fisher Scientific Inc., Waltham, MA, USA).