DNMT3A Antibody (64B1446) - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-80699

Novus Biologicals, part of Bio-Techne

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Chromatin Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), CyTOF-ready, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Whole-Mount, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 64B1446

Format

Azide and BSA Free

Concentration

1 mg/ml

View all DNMT3A Antibodies »

Product Specifications

Immunogen

This DNMT3A Antibody (64B1446) was raised against bacteria expressed recombinant mouse Dnmt3a. The epitope was found to lie near the C-terminus (a.a. 705-908), see Chen et (2002) for details.

Localization

Nuclear

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for DNMT3A Antibody (64B1446) - Azide and BSA Free

Western Blot: DNMT3A Antibody (64B1446)Azide and BSA Free [NBP2-80699]

Western Blot: DNMT3A Antibody (64B1446)Azide and BSA Free [NBP2-80699]

Western Blot: DNMT3A Antibody (64B1446) - Azide and BSA Free [NBP2-80699] - NTERA-2 cells were treated with Zebularine as indicated for 24 hours. Cell lysates were prepared and separated on a 7.5% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was probed with 2 ug/mL anti-Dnmt3a in 1% milk, and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Note the decrease in Dnmt3a expression upon treatment with 500 uM Zebularine (arrowhead). Additional bands at 90 and 75 kDa can also be detected with this antibody and may represent alternative splice variants. Tubulin is shown as a loading control. Image from the standard format of this antibody.
Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - Azide and BSA Free [NBP2-80699]

Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - Azide and BSA Free [NBP2-80699]

Immunocytochemistry/Immunofluorescence: DNMT3A Antibody (64B1446) - Azide and BSA Free [NBP2-80699] - Ntera2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-DNMT3A (68B1446) [NB120-13888] at a 1:200 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.
Immunohistochemistry: DNMT3A Antibody (64B1446) - Azide and BSA Free [NBP2-80699]

Immunohistochemistry: DNMT3A Antibody (64B1446) - Azide and BSA Free [NBP2-80699]

Immunohistochemistry: DNMT3A Antibody (64B1446) - Azide and BSA Free [NBP2-80699] - Detection of DNMT3A on human hepatocellular carcinoma tissue section (NBP2-30221) using 1:100 dilution of DNMT3A antibody (clone 64B1446). The antibody generated a specific cytoplasmic staining in all the cancer cells while some of the cells depicted nucl
View 9 more images

Applications for DNMT3A Antibody (64B1446) - Azide and BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation

reported in scientific literature (PMID 24623306)

Flow Cytometry

1 ug per million cells

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

reported in multiple pieces of scientific literature

Immunohistochemistry Whole-Mount

reported in scientific literature (PMID 26507142)

Immunohistochemistry-Frozen

reported in scientific literature (PMID 22134929)

Immunohistochemistry-Paraffin

5 ug/ml. Use reported in scientific literature (PMID 22134929)
Application Notes
Western Blot: Detects a band of approximately 120 kDa (predicted molecular weight: 102 kDa).

Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: DNMT3A

Methylation of cytosine residues is essential for mammalian development by regulating gene expression, gene silencing and genomic imprinting (1). This process is controlled by the enzyme DNA (cytosine-5)-methyltransferase 3A (DNMt3a), which is encoded by the DNMT3A gene and has a predicted molecular weight of 102 kDa. DNA methyltransferases including DNMt3a are involved in de novo cytosine methylation and maintenance of embryonic and somatic cells through the transfer of methyl groups to specific CpG structures in DNA (2). Somatic mutations in DNMT3A are responsible for cytogenetically normal acute myeloid leukemia (CN-AML). Hypermethylation of CpG islands in tumor suppressor genes or hypomethylation of bulk genomic DNA are linked with cancer (3,4). Mutations in the DNMT3A gene have also been found to cause DNMT3A overgrowth syndrome and myelodysplastic syndromes.

References

1. Thakur, A., Mackin, S. J., Irwin, R. E., O'Neill, K. M., Pollin, G., & Walsh, C. (2016). Widespread recovery of methylation at gametic imprints in hypomethylated mouse stem cells following rescue with DNMT3A2. Epigenetics Chromatin, 9, 53. doi:10.1186/s13072-016-0104-2

2. Ravichandran, M., Lei, R., Tang, Q., Zhao, Y., Lee, J., Ma, L., . . . Dawlaty, M. M. (2019). Rinf Regulates Pluripotency Network Genes and Tet Enzymes in Embryonic Stem Cells. Cell Rep, 28(8), 1993-2003.e1995. doi:10.1016/j.celrep.2019.07.080

3. Pang, Y., Liu, J., Li, X., Xiao, G., Wang, H., Yang, G., . . . Ren, H. (2018). MYC and DNMT3A-mediated DNA methylation represses microRNA-200b in triple negative breast cancer. J Cell Mol Med, 22(12), 6262-6274. doi:10.1111/jcmm.13916

4. Xia, L., Huang, W., Bellani, M., Seidman, M. M., Wu, K., Fan, D., . . . Baylin, S. B. (2017). CHD4 Has Oncogenic Functions in Initiating and Maintaining Epigenetic Suppression of Multiple Tumor Suppressor Genes. Cancer Cell, 31(5), 653-668.e657. doi:10.1016/j.ccell.2017.04.005

Long Name

DNA [Cytosine-5-]-Methyltransferase 3A

Alternate Names

DNA (cytosine-5-)-methyltransferase 3 alpha, DNA (cytosine-5)-methyltransferase 3A, DNA cytosine methyltransferase 3A2, DNA Methyltransferase 3 Alpha, DNA methyltransferase HsaIIIA, DNA MTase HsaIIIA, Dnmt3a, DNMT3A2, EC 2.1.1.37, M.HsaIIIA, TBRS

Gene Symbol

DNMT3A

Additional DNMT3A Products

Product Documents for DNMT3A Antibody (64B1446) - Azide and BSA Free

Product Datasheets

Certificate of Analysis

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Product Specific Notices for DNMT3A Antibody (64B1446) - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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