New England Biolabs Canada
 
XP Monoclonal Antibody

Product Pathways - Chromatin Regulation / Epigenetics

DNMT1 (D63A6) XP® Rabbit mAb #5032

Item# Description List Price Web Price Qty
5032S DNMT1 (D63A6) XP® Rabbit mAb - 100 µl $410.00
$369.00
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5032T DNMT1 (D63A6) XP® Rabbit mAb - 20 µl $171.00
$153.90
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W Human, Mouse, Rat, Monkey Endogenous 200 Rabbit IgG
IF-IC
F

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry

Homology

Species predicted to react based on 100% sequence homology: Hamster, Bovine, Dog, Guinea Pig, Horse.

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).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS7 cells using DNMT1 (D63A6) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L) F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.


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

  • Zhang, Y. et al. (2012) Nucleic Acids Res 40, 4850-60. Applications: Chromatin IP, Western Blotting.
  • Zhang, Y. and Wang, L. (2011) FEBS Lett 585, 1269-75. Applications: Chromatin IP, Western Blotting.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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