DNMT1 Antibody (60B1220.1) - BSA Free

Images

 
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] - An intracellular stain was performed on HeLa cells with DNMT1 [60B1220.1] Antibody NB100-56519AF488 (blue) and a matched isotype control (orange). Cells were ...read more
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
Immunoprecipitation: DNMT1 Antibody (60B1220.1) [NB100-56519] - The PCNAS228I mutation differentially affects PCNA binding to PIP box-containing proteins. GST-PIP pull down of His-S-PCNAWT or PCNAS228I. Figure shows ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, ShSpecies Glossary
Applications WB, Simple Western, ChIP, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, CyTOF-ready, IHC-FrFl, KD
Clone
60B1220.1
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Format
BSA Free
Concentration
1.0 mg/ml

Order Details

DNMT1 Antibody (60B1220.1) - BSA Free Summary

Immunogen
This antibody was raised against a synthetic peptide corresponding to amino acids 637-650 (EKDDREDKENAFKR) of human Dnmt1 (Genbank Accession No. NP_001370).
Isotype
IgG1 Kappa
Clonality
Monoclonal
Host
Mouse
Gene
DNMT1
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Chromatin Immunoprecipitation 1:20 - 1:1000
  • CyTOF-ready
  • Flow (Intracellular)
  • Flow Cytometry 1 - 5 ug/mL
  • Immunocytochemistry/Immunofluorescence 1:10
  • Immunohistochemistry Free-Floating 1:500
  • Immunohistochemistry 1:50-1:500
  • Immunohistochemistry-Frozen 1:500
  • Immunohistochemistry-Paraffin 1:50-1:500
  • Immunoprecipitation 1:10-1:500
  • Knockdown Validated
  • Simple Western 1:100
  • Western Blot 0.1-0.5 ug/ml
Application Notes
Use in ICC/IF reported in scientific literature (PMID 24386225). Use in Flow-intracellular reported in scientific literature (PMID: 24578214). In Simple Western only 10-15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. Knockdown validation (PMID: 31799621).
Reviewed Applications
Read 4 Reviews rated 4.5
using
NB100-56519 in the following applications:

Publications
Read Publications using
NB100-56519 in the following applications:

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 19924110). Porcine reactivity reported in scientific literature (PMID: 23808878). Sheep reactivity reported in scientific literature (PMID: 20413592). Bovine reactivity reported in scientific literature (PMID:27384909).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for DNMT1 Antibody (60B1220.1) - BSA Free

  • AIM
  • CXXC9
  • 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
  • DNMT
  • DNMT1
  • EC 2.1.1.37
  • FLJ16293
  • m.HsaI
  • MCMT
  • MCMTMGC104992

Background

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.

Limitations

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.

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Supplier Logo

Publications for DNMT1 Antibody (NB100-56519)(135)

We have publications tested in 8 confirmed species: Human, Mouse, Rat, Bovine, IHC, Porcine, Sheep, WB.

We have publications tested in 12 applications: ChIP, Chemotaxis, FLOW, Flow Cytometry Control, ICC/IF, IF/IHC, IHC, IHC-Fr, IHC-P, IP, KD, WB.


Filter By Application
ChIP
(1)
Chemotaxis
(17)
FLOW
(1)
Flow Cytometry Control
(1)
ICC/IF
(8)
IF/IHC
(3)
IHC
(1)
IHC-Fr
(1)
IHC-P
(5)
IP
(8)
KD
(1)
WB
(54)
All Applications
Filter By Species
Human
(45)
Mouse
(28)
Rat
(4)
Bovine
(1)
IHC
(1)
Porcine
(1)
Sheep
(1)
WB
(1)
All Species
Showing Publications 1 - 10 of 135. Show All 135 Publications.
Publications using NB100-56519 Applications Species
da C. Fernandes C, da Silva R, de Almeida G et al. Epigenetic Differences Arise in Endothelial Cells Responding to Cobalt-Chromium Journal of Functional Biomaterials 2023-02-26 [PMID: 36976051] (WB, WB) WB WB
Mahadevan KK, McAndrews KM, LeBleu VS et al. Oncogenic Kras G12D specific non-covalent inhibitor reprograms tumor microenvironment to prevent and reverse early pre-neoplastic pancreatic lesions and in combination with immunotherapy regresses advanced PDAC in a CD8 + T cells dependent manner bioRxiv : the preprint server for biology 2023-02-18 [PMID: 36824971] (ChIP, Mouse) ChIP Mouse
Subramani P, Nagarajan N, Mariaraj S, Vilwanathan R Knockdown of sirtuin6 positively regulates acetylation of DNMT1 to inhibit NOTCH signaling pathway in non-small cell lung cancer cell lines Cellular signalling 2023-02-20 [PMID: 36813148] (IP, ICC/IF, WB, Human) IP, ICC/IF, WB Human
Varshney R, Das S, Trahan GD et al. Neonatal intake of Omega-3 fatty acids enhances lipid oxidation in adipocyte precursors iScience 2023-01-20 [PMID: 36590177]
Varshney R, Das S, Trahan GD et al. Neonatal intake of Omega-3 fatty acids enhances lipid oxidation in adipocyte precursors iScience 2023-01-20 [PMID: 36590177]
Dai Z, Liu X, Zeng H, Chen Y Long noncoding RNA HOTAIR facilitates pulmonary vascular endothelial cell apoptosis via DNMT1 mediated hypermethylation of Bcl-2 promoter in COPD Respiratory research 2022-12-17 [PMID: 36527094] (WB, Human) WB Human
Sheth VG, Sharma N, Kabeer SW, Tikoo K Lactobacillus rhamnosus supplementation ameliorates high fat diet-induced epigenetic alterations and prevents its intergenerational inheritance Life sciences 2022-12-15 [PMID: 36343744] (WB, Rat) WB Rat
Zeng W, Jiang H, Wang Y et al. TCF3 Induces DNMT1 Expression to Regulate Wnt Signaling Pathway in Glioma Neurotoxicity research 2022-04-21 [PMID: 35446002] (IP) IP
Liu X, Cui H The palliative effects of folic acid on retinal microvessels in diabetic retinopathy via regulating the metabolism of DNA methylation and hydroxymethylation Bioengineered 2021-12-01 [PMID: 34874218] (Chemotaxis, Rat) Chemotaxis Rat
Kawai T, Richards JS, Shimada M Large-scale DNA demethylation occurs in proliferating ovarian granulosa cells during mouse follicular development Communications biology 2021-11-25 [PMID: 34824385] (FLOW, Mouse) FLOW Mouse
Show All 135 Publications.

Reviews for DNMT1 Antibody (NB100-56519) (4) 4.54

Average Rating: 4.5
(Based on 4 reviews)
We have 4 reviews tested in 1 species: Human.

Reviews using NB100-56519:
Filter by Applications
WB
(2)
ChIP
(1)
IP
(1)
All Applications
Filter by Species
Human
(4)
All Species
Images Ratings Applications Species Date Details
Western Blot DNMT1 NB100-56519
Enlarge
5
reviewed by:
Yichi Xu
WB Human 01/30/2019
View

Summary

ApplicationWestern Blot
Sample Testedcell
SpeciesHuman
LotA-3
Chromatin Immunoprecipitation DNMT1 NB100-56519
Enlarge
5
reviewed by:
Verified Customer
ChIP Human 01/30/2019
View

Summary

ApplicationChromatin Immunoprecipitation
Sample Testedcell
SpeciesHuman
LotA-3

Comments

CommentsMore than 1×10e6 cells and 2.4 μg antibody are optimal.
Western Blot DNMT1 NB100-56519
Enlarge
4
reviewed by:
Douglas Pung
WB Human 04/14/2016
View

Summary

ApplicationWestern Blot
Sample TestedProstate cancer cell lines, LnCAP
SpeciesHuman
  4
reviewed by:
cynthia morimoto
IP Human 12/12/2014
View

Summary

ApplicationImmunoprecipitation
Sample TestedSee PMID 24163373
SpeciesHuman

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Webinar
ChIP Video Protocol
ICC/IF Video Protocol

FAQs for DNMT1 Antibody (NB100-56519). (Showing 1 - 2 of 2 FAQs).

  1. 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.
  2. 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.

Secondary Antibodies

 

Isotype Controls

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Blogs on DNMT1.

The role of DNMT3B in the co-incidence of methyltransferase and tumor suppressor expression in malignancies
Epigenetics is the process of heritable change in gene activity despite alteration of the hosts DNA sequence, essentially causing a change in a phenotype without a change in the genotype of a host. To change the gene sequence without interfering w...  Read full blog post.

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.

Controlling Epigenetic Signaling with Dnmt1 and Dnmt3b
Dnmt1 belongs to the C5-methyltransferase family that repairs cytosines in dsDNA using a nucleophilic attack mechanism. Dnmt1 is the most abundant mammalian DNA methyltransferase. It is the key methylation maintenance enzyme for both DNA replication/r...  Read full blog post.

PCNA Antibodies: Marking Cell Proliferation & DNA Replication
Proliferating Cell Nuclear Antigen (PCNA), also known as the polymerase delta auxiliary protein, is a nuclear protein essential for DNA replication as well as DNA excision and mismatch repair pathways. It has a large role in cell cycle regulation and ...  Read full blog post.

DNMT's: An Overview of 3 DNA Methyltransferases
DNA methyltransferases catalyze the transfer of the methyl group from S-andenosyl methionine (SAM) to DNA. Such methylation has wide ranging function in the cell, including organismal development and cell differentiation. In cancer, abnormal hypermeth...  Read full blog post.

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Recent Reviews

4.5
5
2
4
2
3
0
2
0
1
0

Yichi Xu
01/30/2019
Application: WB
Species: Human

Verified Customer
01/30/2019
Application: ChIP
Species: Human

Douglas Pung
04/14/2016
Application: WB
Species: Human

Bioinformatics

Gene Symbol DNMT1
Entrez
Uniprot