Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Chromatin Regulation / Epigenetics

DNMT1 (D63A6) XP® Rabbit mAb #5032

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC H M R Mk (Hm) (B) (Dg) (GP) (Hr) Endogenous 200 Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Dg=Dog  GP=Guinea Pig  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

DNMT1 (D63A6) XP® Rabbit mAb detects endogenous levels of total DNMT1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of human DNMT1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 and F9 cells using DNMT1 (D63A6) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS cells using DNMT1 (D63A6) XP® Rabbit mAb (green). Actin filaments were labeled using DY-554 phalloidin (red).

Background

Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2 and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as de novo methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither de novo nor maintenance DNA methylation. DNMT1, DNMT3A and DNMT3B together form a protein complex that interacts with histone deacetylases (HDAC1, HDAC2, Sin3A), transcriptional repressor proteins (RB, TAZ-1) and heterochromatin proteins (HP1, SUV39H1), to maintain proper levels of DNA methylation and facilitate gene silencing (3-8). Improper DNA methylation contributes to diseased states such as cancer (1,2). Hypermethylation of promoter CpG islands within tumor suppressor genes correlates with gene silencing and the development of cancer. In addition, hypomethylation of bulk genomic DNA correlates with and may contribute to the onset of cancer. DNMT1, DNMT3A and DNMT3B are over-expressed in many cancers, including acute and chronic myelogenous leukemias, in addition to colon, breast and stomach carcinomas (9-12).

  1. Hermann, A. et al. (2004) Cell. Mol. Life Sci. 61, 2571-2587.
  2. Turek-Plewa, J. and Jagodziński, P.P. (2005) Cell. Mol. Biol. Lett. 10, 631-647.
  3. Kim, G.D. et al. (2002) EMBO J. 21, 4183-4195.
  4. Fuks, F. et al. (2001) EMBO J. 20, 2536-2544.
  5. Geiman, T.M. et al. (2004) Biochem. Biophys. Res. Commun. 318, 544-555.
  6. Rountree, M.R. et al. (2000) Nat. Genet. 25, 269-277.
  7. Pradhan, S. and Kim, G.D. (2002) EMBO J. 21, 779-788.
  8. Fuks, F. et al. (2003) Nucleic Acids Res. 31, 2305-2312.
  9. Mizuno, S. et al. (2001) Blood 97, 1172-1179.
  10. Robertson, K.D. et al. (1999) Nucleic Acids Res. 27, 2291-2298.
  11. Xie, S. et al. (1999) Gene 236, 87-95.
  12. Kanai, Y. et al. (2001) Int. J. Cancer 91, 205-212.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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