Methylation of DNA at cytosine residues plays an important role in regulation of gene expression, genomic imprinting and is essential for mammalian development. Hypermethylation of CpG islands in tumor suppressor genes may be linked with development of cancer. Three families of mammalian DNA methyltransferase genes have been identified which include Dnmt1, Dnmt2 and Dnmt3. Dnmt1 is constitutively expressed in proliferating cells and inactivation of Dnmt1 causes global demethylation of genomic DNA and embryonic lethality. Dnmt1 co-purifies with the retinoblastoma (Rb) tumor suppressor gene product, E2F1, and HDAC1. Dnmt1 also cooperates with Rb to repress transcription from promoters containing E2F binding sites suggesting a link between DNA methylation, histone deacetylase and sequence-specific DNA binding activity, as well as a growth-regulatory pathway that is disrupted in nearly all cancer cells. Phosphorylation of Ser714 is induced by EGF, but the role of this modification is unknown.
Applications:
Suitable for use in ELISA, Western Blot, and Dot Blot. Other applications not tested.
Recommended Dilution:
ELISA: 1:1000
Western Blot: 1:250
Dot Blot: 1:500
Optimal dilutions to be determined by the researcher.
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.