DNMT3A Antibody (64B1446) - BSA Free

Novus Biologicals | Catalog # NB120-13888

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

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

Cited:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Whole-Mount, Western Blot, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Chemotaxis, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 64B1446

Format

BSA Free
View all DNMT3A Primary 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.

Reactivity Notes

Use in Rat reported in scientific literature (PMID:34874218).

Localization

Nuclear

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

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

Western Blot: DNMT3A Antibody (64B1446)BSA Free [NB120-13888]

Western Blot: DNMT3A Antibody (64B1446)BSA Free [NB120-13888]

Western Blot: DNMT3A Antibody (64B1446) [NB120-13888] - Analysis of (A) Dnmt3a transfected 293 cell lysate and (B) untransfected 293 cell lysate using Dnmt3a antibody at 1 ug/mL.
Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Immunocytochemistry/Immunofluorescence: DNMT3A Antibody (64B1446) [NB120-13888] - 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.
Immunohistochemistry-Paraffin: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Immunohistochemistry-Paraffin: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Immunohistochemistry-Paraffin: DNMT3A Antibody (64B1446) [NB120-13888] - 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 nuclear staining also.
Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Immunocytochemistry/Immunofluorescence: DNMT3A Antibody (64B1446) [NB120-13888] - Expression of Dnmt3a in the nuclei of post-mitotic neurons in the olfactory epithelium (OE). DNMT3a (red) is co-expressed with neuron-specific tubulin (green) throughout the development of the olfactory epithelium. Data courtesy of A. Jane Roskams, University of British Columbia.
Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and HEK-293 (B) cells using Dnmt3a antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.
Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - Using the PE direct conjugate, an intracellular stain was performed on NTERA-2 cells with DNMT3A (64B1446) NB120-13888PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.
Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

DNMT3A-Antibody-64B1446-Immunocytochemistry-Immunofluorescence-NB120-13888-img0028.jpg
Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on Jurkat cells with DNMT3A (64B1446) NB120-13888AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
Western Blot: DNMT3A Antibody (64B1446)BSA Free [NB120-13888]

Western Blot: DNMT3A Antibody (64B1446)BSA Free [NB120-13888]

Western Blot: DNMT3A Antibody (64B1446) [NB120-13888] - 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.
Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HepG2 cells with DNMT3A (64B1446) NB120-13888APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).
Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HepG2 cells with DNMT3A (64B1446) NB120-13888AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888]

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HeLa cells with DNMT3A (64B1446) NB120-13888APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).
DNMT3A Antibody (64B1446) - BSA Free

Western Blot: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888] -

nb120-13888_mouse-monoclonal-dnmt3a-antibody-64b1446-20920231721153.jpg
DNMT3A Antibody (64B1446) - BSA Free

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888] -

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888] - Protein levels of Dnmt3a & MeCP2 are reduced in inhibitory neurons of conditional knockout mice.(A) Western blot of half brain hemisphere from Dnmt3a or Mecp2 cKO mice demonstrating loss of each protein. (B) Immunofluorescence (IF) images of WT, Dnmt3a cKO or Mecp2 cKO mice probing for Dnmt3a or MeCP2 (red), the Sun1-sfGFP-myc-PA fusion protein that marks the nuclear envelope & is dependent on Cre expression (green), & DAPI to mark genomic DNA (blue). n = 3 mice per genotype (western), n = 3 mice (IF) representative images for two brain regions shown. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32159514), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
DNMT3A Antibody (64B1446) - BSA Free

Immunocytochemistry/ Immunofluorescence: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888] -

DNMT1 reveals a SCMC-like cortical localization with aberrant localization in Nlrp2tm1a/tm1a derived oocytes.(A) Whole mount immunofluorescence co-staining with anti-DNMT1 and DAPI on the same Nlrp2M+/Z+ embryos used to stain NLRP2 reveals a SCMC-like cortical localization of DNMT1 with a diffuse nuclear stain in later stage embryos. Scale bar represented is 200 μm. (B) DNMT1 staining in paraffin embedded oocyte sections reveals a less cortical, more cytoplasmic intense stain with nucleolar focus in oocytes from Nlrp2tm1a/tm1a dams compared to oocytes from Nlrp2+/+ dams. (C) Whole mount immunofluorescence for DNMT3A in unfertilized control oocytes reveals a characteristic metaphase associated localization and no difference is noted in oocytes derived from Nlrp2tm1a/tm1adams. (D) Whole mount immunofluorescence in unfertilized oocytes reveals that as expected, DNMT3B is not yet expressed in unfertilized control oocytes and no expression is noted in oocytes derived from Nlrp2tm1a/tm1adams. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28317850), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for DNMT3A Antibody (64B1446) - 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)

Knockdown Validated

Validated for Knockdown from CiteAb
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.

Reviewed Applications

Read 3 reviews rated 4.3 using NB120-13888 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 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, 64B1446 clone

Gene Symbol

DNMT3A

Additional DNMT3A Products

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

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Product Specific Notices for DNMT3A Antibody (64B1446) - 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.

Related Research Areas

Adult Neurogenesis
Epigenetics

Citations for DNMT3A Antibody (64B1446) - BSA Free

Customer Reviews for DNMT3A Antibody (64B1446) - BSA Free (3)

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  • DNMT3A Antibody (64B1446)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: cell
    Species: Human
    Verified Customer | Posted 01/30/2019
    DNMT3A Antibody (64B1446) - BSA Free NB120-13888
  • DNMT3A Antibody (64B1446)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: recombinant purified protein
    Species: Human
    Verified Customer | Posted 01/23/2017
    DNMT3A protein
    1ug of recombinant DNMT3A protein. 4-12% Bis tris gel
    DNMT3A Antibody (64B1446) - BSA Free NB120-13888
  • Name: Nathan Burns
    Application: Western Blot
    Sample Tested:
    Species: Other
    Verified Customer | Posted 08/02/2016
    DNMT3A Antibody (64B1446) - BSA Free NB120-13888

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FAQs for DNMT3A Antibody (64B1446) - BSA Free

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  • Q: Do you have any information about the epitope of your antibody, so I could check the chances for the antibody working in the honey bee?

    A:

    This antibody 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. We haven't mapped the binding binding epitope although Chen et (2002) mapped it to a range between 705-908 amino acids: I am not sure what technique you are planning with the antibody, but if you are doing a western blot, we do have a western blot postive control ready to use lysate NBP2-25063 and I would l highly suggest using that.

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