Anti Dnmt3b polyclonal antibody


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Background

Dnmt3b (DNA (cytosine-5-)-methyltransferase 3b) has a role in the establishment and regulation of tissue-specific patterns of methylated cytosine residues (epigenetics). Dnmt3b is thought to function in de novo methylation, rather than maintenance methylation. The protein localizes primarily to the nucleus and its expression is developmentally regulated. Mutations in this gene cause the immunodeficiency-centromeric instability-facial anomalies (ICF) syndrome.
This antibody was prepared and characterized by Prof. S. Tajima of Osaka Univ. and used in Ref.2~4.

Application

1) Western blotting (1,000-fold dilution)
2) Immunoprecipitation (1,000-fold dilution), useful for ChiP assays
3) Indirect immuno-fluorescence staining (1,000-fold dilution)
4) Immunohistochemistry (1000-fold)

<Western blotting>
BAM_70205_01.jpg
The amounts of Dnmt3b in mouse embryos at the stages of E7.0-E16.5 were examined by Western blotting. The embryos were solubilized by sonication or homogenization in the presence of 0.1% SDS. The dissected embryo (10 ug protein) at each stage was subjected to Western blotting with this antibody. Dnmt3b was highly expressed in E7.0 embryo but decreased thereafter and was below the detection level after E10.5. cDNAs of Dnmt3b isoforms (3b1, 3b2, and 3b3) were transiently expressed in 293T cells and these isoforms were also detectable with this antibody.

<Immuno-fluorescence staining>
BAM_70205_02.jpg

Mouse embryos at E4.5, 5.5 and 7.0 were fixed in cold acetone and stained with this antibody or DAPI (blue). The antibody was detected with anti-rabbit IgG conjugated with ALEXA568 (red). The ICM (the inner cell mass) and trophectoderm (T) are indicated by arrows and arrowheads, respectively. The epiblast (E), and the embryonic ectoderm (EE) and extraembryonic region (EX) are indicated by square brackets. Dnmt3b existed at E4.5-7.0, in ICM at E4.5, the epiblast at E5.5, and the embryonic ectoderm at E7.0. 

 

 

 

 

<Reference>
1. Rhee, I. et al. (2002) “DNMT1 and DNMT3b cooperate to silence genes in human cancer Nature 416: 552-6 PMID: 11932749
2. Aoki,A. et al. (2001) “Enzymatic properties of de novo-type mouse DNA (cytosine-methyltransferases.” Nucleic Acids Research 29, 3506-3512 PMID: 11522819
3. Watanabe,D. et al. “Stage- and cell-specific expression of Dnmt3a and Dnmt3b embryogenesis.” Mechanisms of Development 118, 187-190 PMID: 12351185
4. Sakai,Y. et al. (2004) “Co-expression of de novo DNA methyltransferases Dnmt3a2 and Dnmt3L gonocytes of mouse embryos.” Gene Expression Patterns 5, 231-237 PMID: 15567719 

Product List

Product Name Cat# Quantity Price

Anti Dnmt3b, Mouse (Rabbit) Unlabeled DataSheet

BAM-70-205-EX

20 UG N/A

Anti Dnmt3b, Mouse (Rabbit) Unlabeled DataSheet

BAM-70-206-EX

100 UG ¥35,000
$467

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