PRODUCT CODE: EM1901-83

DNMT1 Monoclonal Antibody (EM1901-83)

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

Western blot analysis of DNMT1 on 293 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-83, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of DNMT1 on 293 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-83, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-DNMT1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-83, 1/200) for 30 minutes 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.
  • Flow cytometric analysis of DNMT1 was done on JAR cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-83, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of DNMT1 on 293 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-83, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

DNMT1 Monoclonal Antibody (EM1901-83)

Immunogen

Synthetic peptide.

Host

Mouse

Positive Control

293 cell lysates, human tonsil tissue, JAR.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

A3A8

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein G affinity purified.

MOLECULAR WEIGHT

183 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB:1:500-1:2,000

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

DNMT1

SYNONYMS

ADCADN antibody; AIM antibody; CXXC finger protein 9 antibody; CXXC-type zinc finger protein 9 antibody; CXXC9 antibody; DNA (cytosine 5 ) methyltransferase 1 antibody; DNA (cytosine-5)-methyltransferase 1 antibody; DNA methyltransferase 1 antibody; DNA methyltransferase HsaI antibody; DNA methyltransferase M.HsaI. antibody; DNA MTase antibody; DNA MTase HsaI antibody; DNMT 1 antibody; DNMT antibody; Dnmt1 antibody; DNMT1_HUMAN antibody; Dnmt1o antibody; FLJ16293 antibody; HSN1E antibody; M.HsaI antibody; MCMT antibody; Met1 antibody; MGC104992 antibody; mMmul antibody; MommeD2 antibody

SEQUENCE SIMILARITIES

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

TISSUE SPECIFICITY

Ubiquitous; highly expressed in fetal tissues, heart, kidney, placenta, peripheral blood mononuclear cells, and expressed at lower levels in spleen, lung, brain, small intestine, colon, liver, and skeletal muscle. Isoform 2 is less expressed than isoform 1.

POST-TRANSLATIONAL MODIFICATION

Sumoylated; sumoylation increases activity.; Acetylation on multiple lysines, mainly by KAT2B/PCAF, regulates cell cycle G(2)/M transition. Deacetylation of Lys-1349 and Lys-1415 by SIRT1 increases methyltransferase activity.; Phosphorylation of Ser-154 by CDKs is important for enzymatic activity and protein stability. Phosphorylation of Ser-143 by AKT1 prevents methylation by SETD7 therebye increasing DNMT1 stability.; Methylation at Lys-142 by SETD7 promotes DNMT1 proteasomal degradation.; Ubiquitinated by UHRF1; interaction with USP7 counteracts ubiquitination by UHRF1 by promoting deubiquitination and preventing degradation by the proteasome.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

Methylates CpG residues. Preferentially methylates hemimethylated DNA. Associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand, that is essential for epigenetic inheritance. Associates with chromatin during G2 and M phases to maintain DNA methylation independently of replication. It is responsible for maintaining methylation patterns established in development. DNA methylation is coordinated with methylation of histones. Mediates transcriptional repression by direct binding to HDAC2. In association with DNMT3B and via the recruitment of CTCFL/BORIS, involved in activation of BAG1 gene expression by modulating dimethylation of promoter histone H3 at H3K4 and H3K9. Probably forms a corepressor complex required for activated KRAS-mediated promoter hypermethylation and transcriptional silencing of tumor suppressor genes (TSGs) or other tumor-related genes in colorectal cancer (CRC) cells. Also required to maintain a transcriptionally repressive state of genes in undifferentiated embryonic stem cells (ESCs). Associates at promoter regions of tumor suppressor genes (TSGs) leading to their gene silencing. Promotes tumor growth.