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Dnmt3a Recombinant Rabbit Monoclonal Antibody [ST04-78] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750169

Dnmt3a Recombinant Rabbit Monoclonal Antibody [ST04-78] - BSA and Azide free

Application

  • WB

  • IF-Tissue

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • 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

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

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IF-Tissue

  • 1:1,000

  • IHC-P

  • 1:200-1:3,000

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.

UNIPROT

SwissProt: Q9Y6K1 Human

SwissProt: O88508 Mouse

SwissProt: Q1LZ53 Rat

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

Expand

Images

  • Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/1,000 dilution.<br /><br />Lane 1: C2C12 cell lysate<br />Lane 2: NIH/3T3 cell lysate<br />Lane 3: Mouse brain tissue lysate<br />Lane 4: Mouse skeletal muscle tissue lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 102 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (HA750169) 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 (HA750169) 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.

  • Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/1,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate<br />Lane 2: Hela cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 102 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 2 minutes;<br /><br />8% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (HA750169) 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 (HA750169) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/2,000 dilution.<br /><br />Lane 1: HCT116-WT cell lysate<br />Lane 2: HCT116-KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 102 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 2 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Dnmt3a on different lysates with Rabbit anti-Dnmt3a antibody (HA750169) 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 (HA750169) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Dnmt3a antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/3,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Dnmt3a antibody (HA750169) 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 (HA750169) 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.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Dnmt3a antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>) 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.

    Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Dnmt3a antibody (HA750169) 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 (HA750169) 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.

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Mouse<br />Tissue: Testis<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br />Primary antibody: <a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>, 1/1,000, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1.5 hours at room temperature.

    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: HA750169, 1/1,000, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Rat<br />Tissue: Testis<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br />Primary antibody: <a href="/products/HA750169" style="font-weight: bold;text-decoration: underline;">HA750169</a>, 1/1,000, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1.5 hours at room temperature.

    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: HA750169, 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"