Kit 9999

Immunohistochemistry was performed using the Double-Staining Methods based on Streptavidin Alkaline Phosphatase and Streptavidin Peroxidase labeling techniques (KIT-9999, MXB-China). All of the procedures were carried out according to the manufacturer's instructions. Sections were stained with anti-rabbit CD44 antibody (1:100, ZA-0537, ZSGB, China) and anti-mouse CD24 antibody (1:100, ab31622, Abcam, USA). To control the reliability of the CD44 and CD24 double-staining, single staining with CD44 and CD24 was done. Immunohistochemistry for AR (1:200, ZA-0554, ZSGB, China) and Ki67 (working solution, ZA-0502, ZSGB, China) were carried out as previously reported [36 (link)]. Sections were incubated with goat serum for negative controls of immunoreactions.

The human and monkey tissues were fixed with 4% paraformaldehyde (PFA) and embedded with paraffin. For antigen retrieval, the EDTA retrieval buffer (pH 9.0) was used. The mouse anti-human CD123 (NCL-L-CD123, Leica Biosystems) was used for comparison in this study. The reagents used for the staining were Polymer HRP Detection System (PV-9002, ZSGB-BIO) for single-staining and DouSP (KIT-9999, MXB) for double-staining. The developing reagents were DAB (ZLI-9018, ZSGB-BIO) and VECTOR BLUE SUBSTRATE KIT (SK-5300, Vector Laboratories, Inc).
Immunohistochemistry images were taken by the Nikon binocular microscope (Eclipse Ci-L, Nikon) and imaging system (DS-Ri2, Nikon).