Dnmt3b Recombinant Rabbit Monoclonal Antibody [SY09-07] - BSA and Azide free
Catalog# HA751851
Dnmt3b Recombinant Rabbit Monoclonal Antibody [SY09-07] - BSA and Azide free
-
WB
-
IF-Cell
-
ChIP
-
Human
-
Rat
-
ET1605-9
含抗保成分
-
unconjugated
Overview
Product Name
Dnmt3b Recombinant Rabbit Monoclonal Antibody [SY09-07] - BSA and Azide free
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
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
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.
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
ExpandImages
-
Western blot analysis of Dnmt3b on NCCIT cell lysates with Rabbit anti-Dnmt3b antibody (HA751851) 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 (HA751851) 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 (HA751851) 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 (HA751851) 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 (HA751851) 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 (HA751851) 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 (HA751851) 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 (HA751851) 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"