Western Blot: DNMT1 Antibody (60B1220.1) [NB100-56519] - Analysis of Dnmt1 in 2102EP (human embryonic carcinoma) in the A) absence and B) presence of immunizing peptide, recombinant human Dnmt1 protein in the C) absence ...read more
Immunocytochemistry/ Immunofluorescence: DNMT1 Antibody (60B1220.1) [NB100-56519] - Dnmt1 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and ...read more
Immunohistochemistry-Paraffin: DNMT1 Antibody (60B1220.1) [NB100-56519] - Tissue section of human hepatocellular carcinoma at 1:50 dilution.
Flow Cytometry: DNMT1 Antibody (60B1220.1) [NB100-56519] - Analysis of PE conjugate of NB100-56519. An intracellular stain was performed on HeLa cells with DNMT1 antibody (60B1220.1) NB100-56519PE (blue) and a matched ...read more
Simple Western: DNMT1 Antibody (60B1220.1) [NB100-56519] - Simple Western lane view shows a specific band for Dnmt1 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 ...read more
Rat reactivity reported in scientific literature (PMID: 19924110). Porcine reactivity reported in scientific literature (PMID: 23808878). Sheep reactivity reported in scientific literature (PMID: 20413592).
Packaging, Storage & Formulations
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
0.05% Sodium Azide
Protein G purified
Alternate Names for DNMT1 Antibody (60B1220.1)
CXXC9CXXC finger protein 9
CXXC-type zinc finger protein 9
DNA (cytosine-5-)-methyltransferase 1
DNA (cytosine-5)-methyltransferase 1
DNA methyltransferase 1
DNA methyltransferase HsaI
DNA MTase HsaI
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. NB100-56519 recognizes Dnmt1, a 1616 amino acid protein according to GenBank no. NP_001370.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Bittencourt D, Lee BH, Gao L. Role of distinct surfaces of the G9a ankyrin repeat domain in histone and DNA methylation during embryonic stem cell self-renewal and differentiation. Epigenetics & Chromatin. 2014 (WB, Mouse)
Details: DNMT1 antibody (clone 60B1220.1) used for WB on whole cell extracts from wild-type (TT2) mouse embryonic stem cells /mESCs, G9a-null mESCs or distinct clonal ESC lines (indicated by #1 and #2) stably expressing wild-type G9a (+WT) or G9a harboring the indicated point mutations in the ANK domain (Figure 1B).
FAQs for DNMT1 Antibody (NB100-56519). (Showing 1 - 2 of 2 FAQs).
Could you please tell me the size of your DNMT1 protein along with Expression System? I am also doing mutant DNMT1 protein expression using the truncated part (650-1602=952 aa), using bacterial expression system (E.coli, BL21(De3). But right now I am getting some problem that, my size of protein is little higher than my accurate size? I use Auto-induction method to express the protein.
As you can see from our testing data, we see bands at around 185-200 kDa. Full details of the samples can be found on the image captions. Dnmt1,UniProt P26358, is also highly phosphorylated and acetylated which can raise the apparent molecular weight to higher than the predicted molecular weight. This is why you are seeing the change in size compared to what you might expect from your truncated protein.
We have recently purchased the above mentioned anti-DNMT1 antibody. However, the product datasheet does not have any exact details on the molecular weight of the antibody and images provided are with very wide range protein marker (near DNMT-1 band 116-200kD).
According to Uniprot #P26358 the theoretical molecular weight of human DNMT1 is 183 kDa for isoform 1, ~185 kDa for isoform 2 and 144 kDa for isoform 3. This antibody is designed to detect all 3 of these isoforms.Please not that this protein is subject to multiple posttranslational modifications which may differ by the species, cell type and disease state and these may affect how the protein runs on WB. In case of the WB image (image 1 of 5) we show several human an mouse cell lines, human recombinant protein, and each blot shows at least 1 band which looks like the predicted molecular weight of the target. I agree that the 116-200 range is rather wide and it is impossible to say that the protein runs exactly at 183kDa, but we are confident this is a specific band as, in addition, it is blocked by the blocking peptide. The entire WB image is displayed to show that there were other protein bands observed in some cell lines, also blocked by the peptide. These may be similar epitopes present at other targets or results of posttranslational processing of the DNMT1 protein in these cell types. Further study would be necessary to determine what exactly are these, but we present this data to make the customers aware that there may be some additional bands on the WB. If you are running a WB and ar enot confident that what you see is your target, we recommend to include some positive (overexpression lysate or treatment that induces the protein expression) or/and negative (KO, siRNA, cell line or tissue known not to express your target protein) controls.
The role of DNMT3A in development Epigenetics is the study of heritable change in gene activity despite alteration of the hosts DNA sequence. Change in gene activity done independently of the DNA sequence is achieved by way of histone and DNA methylation. Gene silencing in DNA ... Read full blog post.
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