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.
Key Product Details
Validated by
Species Reactivity
Applications
Label
Antibody Source
Product Specifications
Immunogen
Ala1381-Ala1587
Accession # P26358
Specificity
Clonality
Host
Isotype
Scientific Data Images for Human DNMT1 Antibody
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 Sheep Anti-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 1 µM 5-azacytidine for 24 hours using Sheep Anti-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 in HeLa Human Cell Line.
DNMT1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human DNMT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6110) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Western Blot Shows Human DNMT1 Specificity by Using Knockout Cell Line.
Western blot shows lysates of HEK293T human embryonic kidney parental cell line and DNMT1 knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-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 230 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Applications for Human DNMT1 Antibody
Immunocytochemistry
Sample: Immersion fixed HCT‑116 human colorectal carcinoma cell line untreated and treated with 1μM 5-azacytidine for 24 hours and immersion fixed HeLa human cervical epithelial carcinoma cell line
Knockout Validated
Western Blot
Sample: Jurkat human acute T cell leukemia cell line, K562 human chronic myelogenous leukemia cell line, and MOLT‑4 human acute lymphoblastic leukemia cell line
Reviewed Applications
Read 1 review rated 5 using AF6110 in the following applications:
Formulation, Preparation, and Storage
Purification
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Shipping
Stability & Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: DNMT1
Long Name
Alternate Names
Gene Symbol
UniProt
Additional DNMT1 Products
Product Documents for Human DNMT1 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human DNMT1 Antibody
For research use only
Related Research Areas
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Application: Western BlotSample Tested: See PMID 23935509Species: HumanVerified Customer | Posted 01/08/2015
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars