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PRODUCT CODE: ET1702-77

Dnmt1 Recombinant Rabbit Monoclonal Antibody [JF09-89] (ET1702-77)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of Dnmt1 on HepG2 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 (ET1702-77, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Dnmt1 on HepG2 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 (ET1702-77, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Dnmt1 in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-77, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Dnmt1 in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-77, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Dnmt1 in 293T cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-77, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded mouse placenta tissue using anti-Dnmt1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-77, 1/50) 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.
Western blot analysis of Dnmt1 on HepG2 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 (ET1702-77, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Dnmt1 Recombinant Rabbit Monoclonal Antibody [JF09-89] (ET1702-77)

Immunogen

Synthetic peptide within human dnmt1 aa+l577:l592 1506-1549 / 1616.

Host

Rabbit

Positive Control

HepG2 cell lysates, Hela, HepG2, 293T, mouse placenta tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JF09-89

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

183 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

P26358

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

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.