Dnmt3a Recombinant Rabbit Monoclonal Antibody [ST04-78] - BSA and Azide free
Catalog# HA750169
Dnmt3a Recombinant Rabbit Monoclonal Antibody [ST04-78] - BSA and Azide free
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WB
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IF-Tissue
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IHC-P
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Dnmt3a Recombinant Rabbit Monoclonal Antibody [ST04-78] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human Dnmt3a aa 600-640.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Tissue, IHC-P
Target Molecular Weight
Predicted band size: 102 kDa
Positive Control
C2C12 cell lysate, NIH/3T3 cell lysate, Mouse brain tissue lysate, Mouse skeletal muscle tissue lysate, SH-SY5Y cell lysate, Hela cell lysate, mouse testis tissue, rat testis tissue.
Conjugation
unconjugated
Clone Number
ST04-78
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | IF-Tissue | |
|---|---|---|---|
| human |
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| mouse |
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| rat |
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Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
Methylation at the 5'-position of cytosine is the only known naturally occurring covalent modification of the mammalian genome. DNA methylation requires the enzymatic activity of DNA 5-cytosine methyltransferase (Dnmt) proteins, which catalyze the transfer of a methyl group from S-adenosyl methionine to the 5'-position of cytosines residing in the dinucleotide CpG motif, and this methylation results in transcriptional repression of the target gene. The Dnmt enzymes are encoded by independent genes. Dnmt1 is the most abundant, and it preferentially methylates hemimethylated DNA and coordinates gene expression during development. Additional mammalian Dnmt proteins include Dnmt2 and Dnmt3. Dnmt2 lacks the large N-terminal regulator domain of Dnmt1, is expressed at substantially lower levels in adult tissues, and is likely involved in methylating newly integrated retroviral DNA. Dnmt3a and Dnmt3b are encoded by two distinct genes, but both are abundantly expressed in embryonic stem cells, where they also methylate CpG motifs on DNA.
Background References
1. Dunn J et al. Flow-dependent epigenetic DNA methylation regulates endothelial gene expression and atherosclerosis. J Clin Invest 124:3187-99 (2014).
2. Jefferson WN et al. Persistently altered epigenetic marks in the mouse uterus after neonatal estrogen exposure. Mol Endocrinol 27:1666-77 (2013).
Sequence Similarity
Belongs to the class I-like SAM-binding methyltransferase superfamily. C5-methyltransferase family.
Tissue Specificity
Highly expressed in fetal tissues, skeletal muscle, heart, peripheral blood mononuclear cells, kidney, and at lower levels in placenta, brain, liver, colon, spleen, small intestine and lung.
Post-translational Modification
Sumoylated; sumoylation disrupts the ability to interact with histone deacetylases (HDAC1 and HDAC2) and repress transcription.
Subcellular Location
Cytoplasm, Nucleus, Chromosome.
Synonyms
DNA (cytosine 5) methyltransferase 3 alpha antibody
DNA (cytosine 5) methyltransferase 3A antibody
DNA (cytosine-5)-methyltransferase 3A antibody
DNA cytosine methyltransferase 3A2 antibody
DNA methyltransferase 3 alpha antibody
DNA methyltransferase 3a antibody
DNA methyltransferase HsaIIIA antibody
DNA MTase HsaIIIA antibody
DNM3A_HUMAN antibody
DNMT 3a antibody
ExpandImages
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This data was developed using ET1609-31, the same antibody clone in a different buffer formulation.
Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (ET1609-31) at 1/1,000 dilution.
Lane 1: C2C12 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: Mouse brain tissue lysate
Lane 4: Mouse skeletal muscle tissue lysate
Lysates/proteins at 30 µg/Lane.
Predicted band size: 102 kDa
Observed band size: 130 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-31) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1609-31, the same antibody clone in a different buffer formulation.
Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (ET1609-31) at 1/1,000 dilution.
Lane 1: SH-SY5Y cell lysate
Lane 2: Hela cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 102 kDa
Observed band size: 130 kDa
Exposure time: 2 minutes;
8% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-31) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
This data was developed using ET1609-31, the same antibody clone in a different buffer formulation.
Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (ET1609-31) at 1/2,000 dilution.
Lane 1: HCT116-WT cell lysate
Lane 2: HCT116-KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 102 kDa
Observed band size: 130 kDa
Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-31) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1609-31, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Dnmt3a antibody (ET1609-31) at 1/3,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-31) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1609-31, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Dnmt3a antibody (ET1609-31) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-31) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1609-31, the same antibody clone in a different buffer formulation.
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Testis
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1609-31, 1/1,000, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
This data was developed using ET1609-31, the same antibody clone in a different buffer formulation.
Application: Immunofluorescence (IF-tissue)
Species: Rat
Tissue: Testis
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1609-31, 1/1,000, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"