|Detection of Human DNMT1 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, K562 human chronic myelogenous leukemia cell line, and MOLT‑4 human acute lymphoblastic leukemia cell line. PVDF Membrane was probed with 1 µg/mL of Human DNMT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6110) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for DNMT1 at approximately 183 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|DNMT1 in HCT‑116 Human Cell Line. DNMT1 was detected in immersion fixed HCT‑116 human colorectal carcinoma cell line untreated (left panels) and treated (right panels) with 1μM 5-azacytidine for 24 hours using Human DNMT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6110) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panels; Catalog # NL010) and counterstained with DAPI (blue, lower panels). Specific staining was localized to nuclei and cytoplasm. Nuclear staining was reduced following treatment with 5-azacytidine. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
DNMT1 (DNA methyltransferase 1; also MCMT and CXXC-type Zn finger protein 9) is a 190 kDa member of the C5-methyltransferase family of enzymes. It is ubiquitous in expression, and appears during the S-phase of the cell cycle. DNMT1 catalyzes the attachment of a methyl group to the C5 position of CpG cytosines using AdoMet/adenosylmethionine as a co factor. This acts to silence select genes during X chromosome inactivation and tumorigenesis. Human DNMT1 is 1616 amino acids (aa) in length. It contains an N-terminal regulatory, homodimerization and target recognition region (aa 1-1100), a Gly-Lys linker segment (aa 1109-1120) and a catalytic domain (aa 1121-1616). It is acetylated, phosphorylated, and SUMOylated. There are multiple splice variants. There is a deletion of aa 1-336, alternative start sites at Met122 and 62 aa upstream of the standard start site, and a 17 aa substitution for Pro149. Over aa 1381-1587, human DNMT1 shares 87% aa identity with mouse DNMT1.
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