Rabbit anti asic1

Western blot analyses with T7–T10 DRGs were performed as described previously.³² Nitrocellulose membranes were probed with primary antibodies: mouse anti‐DNMT1 (1:1000, Abcam), rabbit anti‐DNMT3a (1:1000, Abcam), rabbit anti‐DNMT3b (1:1000, Abcam), rabbit anti‐ASIC1 (1:1000, Alomone Labs), and rabbit anti‐GAPDH (1:1000; Hangzhou Goodhere Biotechnology) at 4°C overnight. Subsequently, the membranes were incubated with anti‐rabbit horseradish peroxidase (HRP)‐conjugated (1:2000, Multi Sciences Biotech Co.) or anti‐mouse HRP‐conjugated secondary antibody (1:4000, Sigma) at room temperature for 1 h. The immunoreactive bands were detected with ECL reagents using Image J software on a gel imaging system (Bio‐Rad).

Western blotting was performed to detect the expression of ASIC1 and ASIC2 in colon‐related segments (T13‐L2) of spinal dorsal horn of CON and PMS offspring rats. The experimental method was followed by our previously published articles.³⁰, ³¹ Primary antibodies included rabbit anti‐ASIC1 (1:1000, Alomone Labs, Jerusalem, Israel), rabbit anti‐ASIC2 (1:1000, Alomone Labs, Jerusalem, Israel), and rabbit anti‐GAPDH (1:1000, Goodhere Biotechnology, Hangzhou, China). The secondary antibodies were goat anti‐rabbit peroxidase‐conjugated secondary antibody (1:2000, Multi Sciences Biotech Co., Hangzhou, China).