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Rabbit anti dnmt3a

Manufactured by Abcam

Rabbit anti-DNMT3a is a primary antibody that recognizes the DNA methyltransferase 3A (DNMT3A) protein. DNMT3A is a key enzyme involved in the establishment and maintenance of DNA methylation patterns during development and differentiation.

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2 protocols using rabbit anti dnmt3a

1

Western Blot Analysis of Epigenetic Regulators in DRGs

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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).
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2

ChIP Assay with Chromatin Immunoprecipitation

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The ChIP assays were conducted using the EZ-Magna ChIP™ G - Chromatin Immunoprecipitation Kit (EMD Millipore) 25 (link). Briefly, the homogenate from the DRG was cross-linked with 1% formaldehyde and terminated by glycine. After centrifugation, the collected pellet was resuspended with nuclear lysis buffer and sonicated to shear the cross-linked DNA. After centrifugation, the supernatant was diluted and respectively subjected to immunoprecipitation overnight at 4°C with protein G magnetic beads plus 3 μg of rabbit anti-DNMT3a (Abcam) or 3 μg of normal rabbit serum. Input was used as a positive control. After incubation, protein G magnetic beads were pelleted with the magnetic separator and washed respectively with low salt buffer, high salt buffer, LiCl buffer and TE buffer. The protein/DNA complexes were eluted and reversed to free DNA. The DNA fragments were purified and identified using PCR/real-time PCR with the primers (Supplementary Table 1).
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