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Dnmt3b Recombinant Rabbit Monoclonal Antibody [SY09-07]

Usd: 385

Specification

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Catalog# ET1605-9

Dnmt3b Recombinant Rabbit Monoclonal Antibody [SY09-07]

Application

  • WB

  • IF-Cell

  • ChIP

Reactivity

  • Human

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Dnmt3b Recombinant Rabbit Monoclonal Antibody [SY09-07]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Dnmt3b aa 61-110 / 853.

Species Reactivity

Human, Rat

Validated Applications

WB, IF-Cell, ChIP

Molecular Weight

Predicted band size: 96 kDa

Positive Control

NCCIT cell lysates, NCCIT, C6.

Conjugation

unconjugated

Clone Number

SY09-07

RRID

AB_3069722

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:50-1:250

  • ChIP

  • Use 2 μg for 25 μg of chromatin.

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. Engal E et al. DNMT3B splicing dysregulation mediated by SMCHD1 loss contributes to DUX4 overexpression and FSHD pathogenesis. Sci Adv. 2024 May

2. Man X et al. DNMT3A and DNMT3B in Breast Tumorigenesis and Potential Therapy. Front Cell Dev Biol. 2022 May

Sequence Similarity

Belongs to the class I-like SAM-binding methyltransferase superfamily. C5-methyltransferase family.

Tissue Specificity

Ubiquitous; highly expressed in fetal liver, heart, kidney, placenta, and at lower levels in spleen, colon, brain, liver, small intestine, lung, peripheral blood mononuclear cells, and skeletal muscle. Isoform 1 is expressed in all tissues except brain, skeletal muscle and PBMC, 3 is ubiquitous, 4 is expressed in all tissues except brain, skeletal muscle, lung and prostate and 5 is detectable only in testis and at very low level in brain and prostate.

Post-translational Modification

Sumoylated.; Citrullinated by PADI4.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q9UBC3 Human

Unigene: 117353 Rat

Synonyms

Cytosine 5methyltransferase 3B antibody

DNA antibody

DNA (cytosine 5) methyltransferase 3 beta antibody

DNA (cytosine 5)-methyltransferase 3B antibody

DNA (cytosine-5)-methyltransferase 3B antibody

DNA methyltransferase HsaIIIB antibody

DNA MTase HsaIIIB antibody

DNM3B_HUMAN antibody

Dnmt3b antibody

EC 2.1.1.37 antibody

Expand

Images

  • Western blot analysis of Dnmt3b on NCCIT cell lysates with Rabbit anti-Dnmt3b antibody (<a href="/products/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</a>) at 1/2,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 96 kDa<br />Observed band size: 81 kDa<br /><br />Exposure time: 1 minute 2 seconds; 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/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</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.

    Western blot analysis of Dnmt3b on NCCIT cell lysates with Rabbit anti-Dnmt3b antibody (ET1605-9) at 1/2,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 96 kDa
    Observed band size: 81 kDa

    Exposure time: 1 minute 2 seconds; 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 (ET1605-9) 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.

  • Immunocytochemistry analysis of NCCIT cells labeling Dnmt3b with Rabbit anti-Dnmt3b antibody (<a href="/products/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</a>) at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Dnmt3b antibody (<a href="/products/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</a>) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NCCIT cells labeling Dnmt3b with Rabbit anti-Dnmt3b antibody (ET1605-9) at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Dnmt3b antibody (ET1605-9) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of C6 cells labeling Dnmt3b with Rabbit anti-Dnmt3b antibody (<a href="/products/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Dnmt3b antibody (<a href="/products/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</a>) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C6 cells labeling Dnmt3b with Rabbit anti-Dnmt3b antibody (ET1605-9) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Dnmt3b antibody (ET1605-9) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells with Dnmt3b (<a href="/products/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells with Dnmt3b (ET1605-9) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

  • Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells with Dnmt3b (<a href="/products/ET1605-9" style="font-weight: bold;text-decoration: underline;">ET1605-9</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells with Dnmt3b (ET1605-9) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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Citation