Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Human

Applications

Knockout Validated, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all DNMT1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human DNMT1
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 antibody by Western Blot.

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 antibody in HCT-116 Human Cell Line by Immunocytochemistry (ICC).

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 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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 Antibody Specificity by Using Knockout Cell Line.

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

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

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

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Entrez Gene IDs

1786 (Human); 13433 (Mouse); 84350 (Rat)

Gene Symbol

DNMT1

UniProt

P26358 (Human)

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.

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Product Specific Notices for Human DNMT1 Antibody

For research use only

Customer Reviews for Human DNMT1 Antibody (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 23935509
    Species: Human
    Verified Customer | Posted 01/08/2015

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Protocols

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