DNMT1 Antibody (Biotin)

SKU: orb148076

Description

Mouse monoclonal to DNMT1 (Biotin). Methylation of DNA at cytosine residues plays an important role in regulation of gene expression, genomic imprinting and is essential for mammalian development. Hypermethylation of CpG islands in tumor suppressor genes or hypomethylation of bulk genomic DNA may be linked with development of cancer. To date, 3 families of mammalian DNA methyltransferase genes have been identified which include Dnmt1, Dnmt2 and Dnmt3. Dnmt1 is constitutively expressed in proliferating cells and inactivation of this gene causes global demethylation of genomic DNA and embryonic lethality. Dnmt2 is expressed at low levels in adult tissues and its inactivation does not affect DNA methylation or maintenance of methylation. The Dnmt3 family members, Dnmt3a and Dnmt3b, are strongly expressed in ES cells but their expression is down regulated in differentiating ES cells and is low in adult somatic tissue. Dnmt1 co-purifies with the retinoblastoma (Rb) tumour suppressor gene product, E2F1, and HDAC1. Dnmt1 also cooperates with Rb to repress transcription from promoters containing E2F-binding sites suggesting a link between DNA methylation, histone deacetylase and sequence-specific DNA binding activity, as well as a growth-regulatory pathway that is disrupted in nearly all cancer cells (1-6)..

Images & Validation

• Tested Applications: ChIP, IHC, IP, WB
• Dilution range: WB (1:1000), IHC (1:1000)
• Reactivity: Fish, Human, Mouse, Zebrafish
• Application Notes:
  2 µg/ml of SMC-200 was sufficient for detection of Dnmt1 in 10 µg of mouse ES cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.

Key Properties

• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG1 Kappa
• Clone No.: 60B1220.1
• Immunogen: Raised against a synthetic peptide corresponding to amino acids 637-650 of human DNMT1
• Target: DNMT1
• Molecular Weight: 180kDa
• Purification: Protein G Purified
• Conjugation: Biotin

Storage & Handling

• Storage: Conjugated antibodies should be stored according to the product label
• Buffer/Preservatives: 136.36mM Ethanolamine, 133.23 mM Chlorides, 9.55mM Phosphates, 9.55mM Sodium Bicarbonate
• Concentration: 1 mg/ml
• Disclaimer: For research use only

Alternative Names

DNA Mtase, DNMT, MCMT, DNA methyltransferase 1, AIM, CXXC9, DNA (cytosine-5)-methyltransferase 1, CXXC-type zinc finger protein 9, DNA methyltransferase HsaI

Immunohistochemistry analysis using Mouse Anti-DNMT1 Monoclonal Antibody, Clone 60B1220.1. Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-DNMT1 Monoclonal Antibody at 1:10000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 μl for 2 minutes at RT. Magnification: 40x. ABC solution 1:1000 dilution, DAB solution 100-200uL/ slide.

Western Blot analysis of Human H1299 cell lysate showing detection of DNMT1 protein using Mouse Anti-DNMT1 Monoclonal Antibody, Clone 60B1220.1. Primary Antibody: Mouse Anti-DNMT1 Monoclonal Antibody at 1:1000.

Immunohistochemistry analysis using Mouse Anti-DNMT1 Monoclonal Antibody, Clone 60B1220.1. Tissue: medullar kidney tissue. Species: Mouse. Primary Antibody: Mouse Anti-DNMT1 Monoclonal Antibody at 1:1000. Secondary Antibody: HRP/DAB Detection System: Biotinylated Goat Anti-Mouse, Streptavidin Peroxidase, DAB Chromogen (brown). Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain.

Western Blot analysis of Human 293T cell lysates showing detection of ~180kDa DNMT1 protein using Mouse Anti-DNMT1 Monoclonal Antibody, Clone 60B1220.1. Lane 1: Molecular Weight Marker. Lane 2: Human 293T Cell lysate. Load: 15 μg. Block: 1.5% BSA in 1X TBST. Primary Antibody: Mouse Anti-DNMT1 Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:4000 for 1 hour at RT. Color Development: ECL solution for 5 minutes at RT. Predicted/Observed Size: ~180kDa. Other Band (s): Lower bands are DNMT1 degradation.

Quick Database Links

Gene Symbol

DNMT1

UniProt

P26358

NCBI Gene ID

1786

RefSeq (Protein):NP_001370