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Product Pathways - Chromatin Regulation / Epigenetics

DNMT3B (D7O7O) Rabbit mAb #67259

Item# Description List Price Web Price Qty
67259S DNMT3B (D7O7O) Rabbit mAb - 100 µl $372.00
$334.80
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67259T DNMT3B (D7O7O) Rabbit mAb - 20 µl $159.00
$143.10
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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
VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W Human Endogenous 96 Rabbit IgG
IP
IF-IC

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry)

Specificity / Sensitivity

DNMT3B (D7O7O) Rabbit mAb recognizes endogenous levels of total DNMT3B protein. This antibody also detects a non-specific protein of approximately 65 kDa in multiple cell lines. Based on sequence homology, this antibody should recognize all isoforms of DNMT3B. This antibody shows low sensitivity in IF-IC, where it only detects DNMT3B in high expressing cells. However, this clone detects DNMT3B in both high and low expressing cells by western blot.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein surrounding Ala395 of human DNMT3B protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HCT 116 DNMT3B wild-type (HCT 116 WT), HCT 116 DNMT3B knockout (HCT 116 KO), K-562, and NCCIT cells, using DNMT3B (D7O7O) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of NCCIT (left) and HEK 293 (right) cells using DNMT3B (D7O7O) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). HEK 293 cells express very low levels of DNMT3B protein.

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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