Mouse anti cd163 antibody

Immunofluorescence staining was used to verify the expressions of CD68 and CD163. The tissue sections of PL6 were deparaffinized and rehydrated, followed by antigen retrieval. After being blocked for 1 h in 3% bovine serum albumin (BSA) at 37°C, the tissue sections were incubated overnight at 4°C with the following primary antibodies: rabbit anti-CD68 antibody (1:100; Abcam, Cambridge, UK) and mouse anti-CD163 antibody (1:100; Abcam). Subsequently, sections were incubated with the secondary antibodies for 1 h at 37°C, followed by counterstaining with DAPI. The tissue sections were then observed and photographed under the inverted microscope.

The tissue blocks sampled from 116 cases of CRC were fixed with 4% formaldehyde, conventionally dehydrated before embedding in paraffin and sliced into 4-µm sections Immunohistochemical of CD163, CD4 and CXCR5 were performed using the EnVision two-step method on the Dako Omnis automated immunohistochemical stainer. All target antigens were prepared with high-pressure sodium citrate buffer at pH 6.0. PBS was used in place of the primary antibody for a blank control, and already-known positive cases were used as positive controls. The specific steps were carried out according to the kit manufacturers instructions. Rabbit anti-CD4 antibody, mouse anti-CD163 antibody and mouse anti-CXCR5 antibody were provided by Abcam. Immunohistochemical sections were scanned by the pathological section scanner (3DHISTECH). Five 400-fold visual fields were randomly selected, and positive cells were counted with Image pro plus 6.0 software to measure the average value.