Antibodies to DNA Methyltransferases (Dnmt Family) - FocusOn 089

Acris Antibodies Panel to the Dnmt Family

Acris Antibodies offers a range of monoclonal and polyclonal antibodies to Dnmt1, Dnmt2, Dnmt3a/3b and Dnmt3-like (Dnmt3L) proteins. All antibodies are suitable for Western blot, ELISA and/or immunoprecipitation / immunohistochemistry (see Product list).

Introduction

In mammals, epigenetic information is established and maintained via the postreplicative methylation of cytosine residues by the DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b.

Dnmt1 is required for maintenance methylation whereas Dnmt3a and Dnmt3b are responsible for de novo methylation. A systematic analysis of interactions among the Dnmt family members has revealed that Dnmt3L interacts with the C-terminal domain of Dnmt3a and Dnmt3b. The interaction between Dnmt3-like (Dnmt3L) and the C-terminal domains of Dnmt3a and Dnmt3b suggests a mechanism whereby the enzymatically inactive Dnmt3L brings about the methylation of its substrate by recruiting an active methylase. Quantitative analysis of genomic cytosine methylation levels demonstrated a robust Dnmt activity for the methyltransferases Dnmt3a and Dnmt3b. In addition, a weak but significant activity for Dnmt2 has been detected. Subsequent methylation tract analysis by genomic bisulphite sequencing reveals that Dnmt3 enzymes preferentially methylate CpG dinucleotides in a processive manner, whereas Dnmt2 methylates isolated cytosine residues in a non-CpG dinucleotide context.

Inappropriate over-expression of Dnmt can lead to inadequate methylation of DNA and thus, misregulation of the cell’s genes. If, for example, inappropriate methylation occurs on tumour suppressor genes, expression of the tumour suppressor genes is turned off, and cancer may develop. The methylation of tumour suppressor genes has been observed to occur in a wide variety of human cancers.

Fig. 1: Western blot analysis of Dnmt1 in 2102EP (human embryonic carcinoma): in A) absence and B) presence of immunizing peptide; recombinant human Dnmt1 protein in the C) absence and D) presence of immunizing peptide; NIH 3T3 (mouse embryonic fribroblast) in the E) absence and F) presence of immunizing peptide; D3 (mouse embryonic stem cell) in the G) absence and H) presence of immunizing peptide using Dnmt1 antibody Cat.-No. SM7026P
Fig. 2: IHC analysis of Dnmt1 using Cat.-No. SM7026P at 2 µg/ml on formalin-fixed, paraffin-embedded (FFPE) medullar kidney tissue sections
Fig. 3: Immunofluorescence analysis of paraformaldehyde-fixed A431, using Dnmt1 antibody Cat.-No. AP22393PU-N at 1/200 dilution (right figure merged with DNA probe)
Fig. 4: Staining of human thymus (FFPE) using Dnmt2 antibody Cat.-No. SM7027P at 5 µg/ml followed by biotinylated secondary antibody, alkaline phosphatase-streptavidin and chromogen
Fig. 5: Immunohistochemistry analysis of Dnmt3b Cat.-No. AP01182PU-N in human liver carcinoma tissue (FFPE)
Fig. 6: Staining of human cancer tissue (FFPE) with Dnmt3L antibody Cat.-No. AP11060PU-N using peroxidase-conjugated secondary antibody followed by DAB staining

Key references

 
Cheng X et al: Mammalian DNA methyltransferases: a structural perspective.
Structure, Mar 2008; 16(3): 341-50.
 
Mund C et al: Comparative analysis of DNA methylation patterns in transgenic drosophila overexpressing mouse DNA methyltransferases.
Biochem J, Mar 2004; 15, 378 (Pt3): 763-8.
 
Margot JB et al: Interactions within the mammalian methyltransferase family.
BMC Mol Biol, May 2003; 30: 4-7.
 
Liu K et al: Endogenous assays of DNA methyltransferases: Evidence for differential activities of Dnmt1, Dnmt2, and Dnmt3 in mammalian cells in vivo.
Mol Cell Biol, Apr 2003; 23(8): 2709-19.
 
Kareta MS et al: Reconstitution and mechanism of the stimulation of de novo methylation by human Dnmt3L.
J Biol Chem, Sep 2006; 8, 281(36): 25893-902.

Further readings

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

DNMT1 (1603-1616) antibody

Figure 1. Staining DNMT1 in Testis by Immunohistochemistry using Formalin-Fixed Paraffin-Embedded (FFPE) tissue. Rabbit Purified Hu E, P
50 µg / €470.00
  Acris Antibodies GmbH

DNMT1 (177-550) antibody

Placenta: Formalin-Fixed Paraffin-Embedded (FFPE) Chicken Ig Azide Free Hu P, WB
50 µg / €470.00
  Acris Antibodies GmbH

DNMT1 antibody

Western Blot analysis of Human DNMT1 expression in nuclear extracts of HeLa cells (50 µg/lane). SP2063P DNMT1 antibody was used at the indicated concentrations. Rabbit Aff - Purified Hu IP, WB
50 µg / €330.00
  Acris Antibodies GmbH

DNMT2 / TRDMT1 antibody

Immunohistochemistry: AP11029PU-N Dnmt2 antibody staining of Formalin-Fixed, Paraffin-Embedded Human cancer tissue. The primary antibody was peroxidase-conjugated to the secondary antibody, followed by DAB staining. Rabbit Ig Purified Hu, Ms P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

DNMT2 / TRDMT1 (N-term) antibody

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu, Ms P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

DNMT3A antibody

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

DNMT3A (N-term) antibody

Formalin-fixed and paraffin-embedded human lung carcinoma tissue reacted with Dnmt3a antibody (N-term) (Cat.#AP11051PU-N), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

DNMT3B antibody

Immunohistochemistry (IHC) analyses of Dnmt3b antibody (Cat.-No. AP01182PU-N) in Paraffin-Embedded Human liver carcinoma tissue. Rabbit Aff - Purified Hu P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

DNMT3B antibody

Formalin-fixed and paraffin-embedded human prostata carcinoma tissue reacted with Dnmt3b antibody (Cat.#AP11052PU-N), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

DNMT3L (322-337) antibody

Figure 1. Staining DNMT3L in Tonsil by Immunohistochemistry using Formalin-Fixed Paraffin-Embedded (FFPE) tissue. Rabbit Purified Hu E, P
50 µg / €470.00
  Acris Antibodies GmbH

DNMT3L (C-term) antibody

AP11060PU-N DNMT3L antibody staining of Formalin-Fixed, Paraffin-Embedded Human cancer tissue using Peroxidase-conjugated secondary antibody, followed by DAB staining. Rabbit Ig Purified Hu P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

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