DNMT3B Antibody [K20B15] | Primary Antibodies

Application: Reactivity: Human

Lane 1: HCT116, Lane 2: HCT116 (KO DNMT3B), Lane 3: K562, Lane 4: NCCIT

Usage Information

Dilution
WB1:1000 IP1:50 IF1:800

Application
WB, IP, IF

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
96 kDa

Positive Control	HCT 116 cells; K-562 cells; NCCIT cells
Negative Control	HEK 293 cells

Datasheet & SDS

Biological Description

Specificity
DNMT3B Antibody [K20B15] detects endogenous levels of total DNMT3B protein.

Clone
K20B15

Synonym(s)
DNA (cytosine-5)-methyltransferase 3B; Dnmt3b; DNA methyltransferase HsaIIIB (DNA MTase HsaIIIB; M.HsaIIIB); DNMT3B

Background
DNMT3B is a critical de novo DNA methyltransferase responsible for establishing methylation patterns during early embryonic development, particularly at implantation, by targeting centromeric, pericentromeric, subtelomeric repeats, and intragenic regions of active genes to regulate gene expression, chromatin structure, and cellular differentiation. DNMT3B contains an N-terminal PWWP domain that binds H3K36me3 and regulates tetramer assembly, an ADD domain that recognizes unmethylated H3K4 tails to autoinhibit the catalytic methyltransferase (MTase) domain through intramolecular interactions while facilitating nucleosome core binding via acidic patch contacts, and a C-terminal MTase domain with a target recognition domain (TRD) that mediates DNA binding, homodimerization, and catalyzes methyl group transfer from S-adenosyl methionine to cytosine C5 at both CpG and non-CpG sites, often in concert with DNMT3A and DNMT3L for processive modification of nucleosomal linker DNA. DNMT3B collaborates with other DNMTs and chromatin factors such as HDACs and HP1 to drive epigenetic silencing, genomic imprinting, and heterochromatin maintenance, with its oligomerization and histone-tail sensing enabling context-specific targeting. The mechanistic interplay between PWWP and ADD domains relieves autoinhibition in response to H3K4me0/H3K36me3, promoting MTase access to DNA, while mutations—such as those seen in ICF syndrome- impair repeat methylation and cause chromosomal instability. Dysregulation of DNMT3B, including overexpression or pathogenic variants, contributes to diseases like ICF syndrome and various cancers by causing aberrant DNA hypermethylation or hypomethylation of tumor suppressor genes and oncogenes.

Background

DNMT3B is a critical de novo DNA methyltransferase responsible for establishing methylation patterns during early embryonic development, particularly at implantation, by targeting centromeric, pericentromeric, subtelomeric repeats, and intragenic regions of active genes to regulate gene expression, chromatin structure, and cellular differentiation. DNMT3B contains an N-terminal PWWP domain that binds H3K36me3 and regulates tetramer assembly, an ADD domain that recognizes unmethylated H3K4 tails to autoinhibit the catalytic methyltransferase (MTase) domain through intramolecular interactions while facilitating nucleosome core binding via acidic patch contacts, and a C-terminal MTase domain with a target recognition domain (TRD) that mediates DNA binding, homodimerization, and catalyzes methyl group transfer from S-adenosyl methionine to cytosine C5 at both CpG and non-CpG sites, often in concert with DNMT3A and DNMT3L for processive modification of nucleosomal linker DNA. DNMT3B collaborates with other DNMTs and chromatin factors such as HDACs and HP1 to drive epigenetic silencing, genomic imprinting, and heterochromatin maintenance, with its oligomerization and histone-tail sensing enabling context-specific targeting. The mechanistic interplay between PWWP and ADD domains relieves autoinhibition in response to H3K4me0/H3K36me3, promoting MTase access to DNA, while mutations—such as those seen in ICF syndrome- impair repeat methylation and cause chromosomal instability. Dysregulation of DNMT3B, including overexpression or pathogenic variants, contributes to diseases like ICF syndrome and various cancers by causing aberrant DNA hypermethylation or hypomethylation of tumor suppressor genes and oncogenes.

References
https://pubmed.ncbi.nlm.nih.gov/15456878/ https://pubmed.ncbi.nlm.nih.gov/37941146/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%