Rabbit anti chga antibody

The slides were incubated overnight at 4°C with the primary antibody, washed in PBS and incubated with secondary antibodies for 1.5 hours at room temperature. This was followed by washing with PBS and mounting with Vectashield‐DAPI mounting medium. Primary antibodies used in our experiment were as follows: mouse anti‐RET antibody (1:100; Santa Cruz, Santa Cruz, CA) and rabbit anti‐ChgA antibody (1:200; Abcam). Secondary antibodies were Alexa Fluor 647‐conjugated anti‐mouse IgG (1:400; Abcam) and Alexa Fluor 488‐conjugated anti‐rabbit IgG (1:400; Abcam), and DAPI (Abcam). Images were taken on Zeiss LSM 880 with an AiryScan confocal laser scanning microscope and were analysed with zen 2009 Light Edition software.

Endoscopic biopsy colon tissues and mice colon were fixed overnight at 4°C in 4% paraformaldehyde and was subsequently embedded in paraffin. Fixed and embedded tissues were sectioned (4 μm) into slides. Immunohistochemical staining of human colon biopsy was performed using rabbit anti‐GDNF (1:500, Abcam, Cambridge, UK), rabbit anti‐ChgA antibody (1:300; Abcam), rat anti‐serotonin antibody (1:500; Abcam) and mouse anti‐RET antibody (1:300; Abcam). Mice colon sections were performed using rat anti‐serotonin antibody (1:500; Abcam).