Alexa fluor 488 donkey anti rabbit igg antibody

Fixed prostatic and bladder tissues from different groups were processed using routine paraffin techniques to prepare sections at a thickness of 4 μm. This was followed by deparaffinization, rehydration and staining with haematoxylin and eosin (H&E).
Immunofluorescent (IF) staining was used to identify the sites of CD45 and Foxp3 expression in mouse prostate tissues. Tissue sections of prostate were stained with primary antibodies CD45 (1:200, Abcam) and Foxp3 (1:200, GeneTex). Then, sections were stained with the fluorescent secondary antibodies Alexa Fluor^® 488 donkey anti‐rabbit IgG antibody (Abcam) and Alexa Fluor^® 594 donkey antimouse IgG antibody (Abcam); and with 4′6‐diamidino‐2‐phenylindole (DAPI, Sigma‐Aldrich Co., Ltd). Sections were then mounted with antifade mounting medium (Abcam).

Frozen sections were rewarmed at 4 °C for 30 min, and 5% goat serum was used to block the slices for 2 h. Then, the sections were incubated with Cy3-labeled GFAP (1:400 dilution; Abcam, Shanghai, China) and Iba1 (1:400 dilution; Wako, Japan)/NF-κB p65 (1:500 dilution; Abcam, Shanghai, China), followed by washing three times with PBS for 10 min each time. Alexa Fluor 488 donkey anti-rabbit IgG antibody (1:1000 dilution; Abcam, Shanghai, China) was added and then the slices were incubated for 2 h in the dark, followed by washing in PBS 3 times for 10 min each, incubation in DAPI for 10 min at room temperature in the dark, and then washing in PBS 3 times at 10 min per time. Fluorescence decay was used to seal the tablets, and the images were collected using a laser scanning confocal microscope (Carl Zeiss I800, Jena, Germany).