Anti-Dnmt3b antibody, affinity-purified (rabbit polyclonal), CHIP grade

$350.00

SKU: 70-206 Research Area: Applications:

Description

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 expressions are 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.

 

Applications

  1. Western blot 1,000-fold dilution
  2. Immunoprecipitation 1,000-fold dilution, useful for ChiP assays
  3. Indirect immunofluorescence staining 1,000-fold dilution
  4. Immunohistochemistry (1,000-fold)

 

Specification

Immunogen: Highly purified recombinant mouse Dnmt3b (amino acids 1-181) (accession no. AF068626

Antibody:  Affinity-purified with the recombinant Dnmt3b

Reactivity: Reacts with mouse Dnmt3b. It may also react with other rodent homologs due the sequence homology, but it does not react with human homolog due to the sequence difference.

Form: 1 mg/ml in PBS, 50% glycerol, 0.05% sodium azide (and trace of ammonium sulfate)

Storage: ship 4°C and store at -20°C or long term storage -80°C

 

Data Link

UniProtKB/Swiss-Prot O88509 (DNM3B_MOUSE)

 

References: This antibody was used for Western blot and immunohistochemistry in Ref. 2~4.

  1. Rhee I et al “DNMT1 and DNMT3b cooperate to silence genes in human cancer cells” Nature 416: 552-6 (2002) PMID: 11932749

2 Aoki A et al “Enzymatic properties of de novo-type mouse DNA (cytosine-5) methyltransferases” Nucleic Acids Research 29: 3506-3512 (2001) PMID: 11522819

  1. Watanabe D et al “Stage- and cell-specific expression of Dnmt3a and Dnmt3b during embryogenesis” Mechanisms of Development 118: 187-190 (2002) PMID: 12351185
  2. Sakai Y et al “Co-expression of de novo DNA methyltransferases Dnmt3a2 and Dnmt3L in gonocytes of mouse embryos” Gene Expression Patterns 5: 231-237 (2004) PMID: 15567719

The data of Western blot and immunofluorescence staining obtained by using this antibody are shown in the next page.

 

Related product: #70-201 Anti-Dnmt1 (1-248) antibody, affinity-purified (rabbit polyclonal)

#70-203 Anti-Dnmt1 (1037-1086) antibody, affinity-purified (rabbit polyclonal)

Fig. 1 Western blot of Dnmt3b.

The amounts of Dnmt3b in mouse embryos at the stages of E7.0-E16.5 were examined by Western blot. The embryos were solubilized by sonication or homogenization in the presence of 0.1% SDS. The dissected embryo (10 µg protein) at each stage was subjected to Western blot 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.

Fig. 2 Expression of Dnmt3b in mouse embryos was examined by immunofluorescence staining.

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 AF-568 (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.

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70-206 Dnmt3b antibody application/pdf 370 KB

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