Anti–DNMT3B polyclonal antibody

Catalog #

25219

$415 *

Size: 50 µg

Qty

Bulk/Custom

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Description

Polyclonal antibody raised in rabbit against mouse DNMT3B (DNA methyltransferase 3B), using 3 KLHconjugated synthetic peptides containing sequences from different parts of the protein.

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Synonyms

DNA (cytosine-5)-methyltransferase 3B, DNMT-3B

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Product Info

Storage and Usage

Citations

Clonality

polyclonal

Host Species/Expression System

Rabbit

Purification

Affinity purified

Format

Aqueous buffer solution

Formulation

PBS containing 0.05% azide and 0.05% ProClin 300

Concentration

50 µg/ 16 µl

Immunogen

synthetic peptide

Background

DNMT3B (UniProtKB/Swiss–Prot entry Q9UBC3) catalyses the genome wide de novo methylation of CpG residues, which regulates gene expression. DNMT3B is essential for development. DNA methylation on CpG residues is coordinated with methylation of histones. Six different isoforms of DNMT3B, produced by alternative splicing, exist although isoforms 4 and 5 may not be functional due to the absence of two conserved methyltransferase motifs.

Storage/Stability

Store at –80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at –20°C for at least one month.

Assay Conditions

Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of antibody directed against DNMT3B (Cat. No. 25219). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:220,000.

Western blot analysis using the antibody directed against DNMT3B
Whole cell extracts (25 μg) from HeLa cells were analysed by Western blot using the antibody against DNMT3B (Cat. No. 25219) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Immunofluorescence using the antibody directed against DNMT3B
Human HeLa cells were stained with the antibody against DNMT3B (Cat. No. 25219) and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the DNMT3B antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Applications

ELISA (1:10,000)
WB (1:1000)
IF (1:1000)

Shipping Temperature

-20°C or-80°C

Species Reactivity

Human, mouse

Notes

The optimal antibody concentration should be determined by the end–user.

Warnings

Avoid freeze/thaw cycles

Scientific Category

Antibodies

Regulatory Status

Research Use Only

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Anti–DNMT3B polyclonal antibody

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