Human DNMT1 Antibody
R&D Systems, part of Bio-Techne | Catalog # AF6110
Key Product Details
Validated by
Knockout/Knockdown, Biological Validation
Species Reactivity
Human
Applications
Immunocytochemistry, Knockout Validated, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
Product Specifications
Immunogen
E. coli-derived recombinant human DNMT1
Ala1381-Ala1587
Accession # P26358
Ala1381-Ala1587
Accession # P26358
Specificity
Detects human DNMT1 in Western blots.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
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.Applications for Human DNMT1 Antibody
Application
Recommended Usage
Immunocytochemistry
1-15 µg/mL
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
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
DNMT1
is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in
DNMT1 knockout HEK293T cell line.
Western Blot
1 µg/mL
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
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
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 1 review rated 5 using AF6110 in the following applications:
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 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.
Background: DNMT1
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.
Long Name
DNA [Cytosine-5-]-Methyltransferase 1
Alternate Names
CXXC9, MCMT
Gene Symbol
DNMT1
UniProt
Additional DNMT1 Products
Product Documents for Human DNMT1 Antibody
Product Specific Notices for Human DNMT1 Antibody
For research use only
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