R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.
DNMT3A (E9P2F) Rabbit mAb #49768
Filter:
- WB
- IP
- ChIP
Supporting Data
| REACTIVITY | H M R Mk |
| SENSITIVITY | Endogenous |
| MW (kDa) | 85, 95, 130 |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- ChIP-Chromatin Immunoprecipitation
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
- Mk-Monkey
Product Information
Product Usage Information
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
Protocol
Specificity / Sensitivity
DNMT3A (E9P2F) Rabbit mAb recognizes endogenous levels of total DNMT3A protein. This antibody detects multiple isoforms of DNMT3A, including isoform 1 and isoform 2. This antibody does not cross-react with DNMT3B or other DNMT proteins.
Species Reactivity:
Human, Mouse, Rat, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human DNMT3A 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 overexpressed in many cancers, including acute and chronic myelogenous leukemias, in addition to colon, breast, and stomach carcinomas (9-12).
- Hermann, A. et al. (2004) Cell. Mol. Life Sci. 61, 2571-87.
- Turek-Plewa, J. and Jagodziński, P.P. (2005) Cell. Mol. Biol. Lett. 10, 631-47.
- Kim, G.D. et al. (2002) EMBO J. 21, 4183-95.
- Fuks, F. et al. (2001) EMBO J. 20, 2536-44.
- Geiman, T.M. et al. (2004) Biochem. Biophys. Res. Commun. 318, 544-55.
- Rountree, M.R. et al. (2000) Nat. Genet. 25, 269-77.
- Pradhan, S. and Kim, G.D. (2002) EMBO J. 21, 779-88.
- Fuks, F. et al. (2003) Nucleic Acids Res. 31, 2305-12.
- Mizuno, S. et al. (2001) Blood 97, 1172-9.
- Robertson, K.D. et al. (1999) Nucleic Acids Res. 27, 2291-8.
- Xie, S. et al. (1999) Gene 236, 87-95.
- Kanai, Y. et al. (2001) Int. J. Cancer 91, 205-12.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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