R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.
DNMT3B (D7O7O) Rabbit mAb #67259
Filter:
- WB
- IP
- IF
Supporting Data
| REACTIVITY | H |
| SENSITIVITY | Endogenous |
| MW (kDa) | 96 |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IF-Immunofluorescence
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunofluorescence (Immunocytochemistry) | 1:1600 |
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
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.
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein surrounding Ala395 of human 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 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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