immunofluorescence of rabbit anti-murine RELMβ by Peprotech

实验概要

The following protocol provides a method of immunofluorescence of rabbit anti-murine RELMβ by Peprotech.

实验步骤

The following protocol used B6 mice that were infected with 200 embryonated Trichuris muris eggs and sacrificed 21 days post-infection. The tip of the cecum was removed, rinsed in PBS, and fixed in 4% PFA. Following fixation, the tissue was embedded in paraffin and cut into 5 μM sections.

1. Deparaffinize and rehydrate the tissue section.

2. Perform heat-induced antigen retrieval by boiling the tissue section in 10mM citrate buffer for 25 minutes.

3. Incubate the tissue section with blocking buffer for 20 minutes.

4. Incubate the tissue section overnight at 4˚C with Rabbit Anti-Murine RELMβ at 0.2 μg/mL in PBS with 0.01% Triton-X and 0.5% BSA. Wash the slide twice for three minutes (1X PBS/0.05% Tween 20).

5. Incubate the tissue section with a fluorescent conjugated secondary antibody for 2 hours at room temperature. Wash the slide twice for three minutes.

6. Counterstain the tissue section with DAPI.