DNA methylation can help to regulate transcriptional silencing via repressive transcription complexes, which include methyl-CpG-binding domain proteins and histone deacetylases (HDACs). DNMT1, the core enzyme for mammalian DNA methylation, can also establish a repressive transcription complex consisting of DNMT1, HDAC2, and a third protein, termed DMAP1 for 'DNMT1-associated protein.' The 467-amino acid DMAP1 protein shares approximately 98% amino acid sequence homology with the mouse protein. DMAP1 interacts directly with the N-terminal region of DNMT1, and DMAP1 can repress transcription independently of histone deacetylase activity. DNMT1, HDAC2, and DMAP1 form a complex in vivo, and DMAP1 can interact directly with the transcriptional corepressor TSG101. The DNMT1-DMAP1 exists throughout the S phase; HDAC2 joins DNMT1 and DMAP1 only during late S phase. This provides a regulated means to deacetylate heterochromatin following replication.
Applications:
Suitable for use in ELISA, Western Blot, Immunohistochemistry, Immunocytochemistry. Other applications not tested.
Recommended Dilution:
ELISA: 1:1000
Western Blot: 1:100-1:500
Immunohistochemistry (paraffin): 1:50-100
Immunocytochemistry: 1:500
Optimal dilutions to be determined by the researcher.
Positive Control:
HL60 cell lysate
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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