Rabbit anti dnmt3b

An affinity-purified rabbit polyclonal antibody to the 67 amino acid VDR readthrough peptide was prepared by Proteintech to generate anti-VDRx. The following commercially available antibodies were also used. Rabbit anti-VDR (Abcam, ab109234), mouse anti-HA (Covance, clone 16B12), mouse anti-β-actin (Sigma, A3853), rabbit anti-DNMT3B (Abcam, ab79822), and rabbit anti-EEF2 (Cell Signaling Technology, 2332).

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).

INS‐1 cells were harvested with ice‐cold lysis buffer supplemented with complete proteinase inhibitor mixture (Roche). Other specific information of Western blotting assay was performed as previously described previously.¹⁹ The antibodies used are as follows: rabbit anti‐CASK antibody (1:1000; Cell Signaling), rabbit anti‐DNMT1 (1:1000; Cell Signaling), mouse anti‐DNMT3a (1:1000; Santa Cruz Biotechnology), rabbit anti‐DNMT3b (1:1000; Abcam), rabbit anti‐Akt(pan) antibody (1:1000; Cell Signaling), rabbit anti‐p‐Akt Ser473 antibody (1:5000; Abcam), rabbit anti‐iNOS Antibody (1:1000; Cell Signaling) or rabbit anti‐β‐tubulin antibody (1:4000; Cell Signaling).