5-hydroxymethylcytosine (5-hmC) Antibody (mouse)
One of the only two monoclonal antibodies raised against 5-hydroxymethylcytosine (5-hmC). 5-hmC is a recently discovered DNA modification which results from the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of oxygenases. Preliminary results indicate that 5-hmC may have important roles distinct from 5-methylcytosine (5-mC). Although its precise role has still to be shown, early evidence suggests a few putative mechanisms that could have big implications in epigenetics.
| Lot | 002 |
|---|---|
| Concentration | 2.07 µg/µl |
| Species reactivity | Human, mouse, other (wide range): positive |
| Type | Monoclonal |
| Purity | Protein A purified monoclonal antibody |
| Host | Mouse |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| hMeDIP | 2 μg/IP | Fig 1 |
| Dot Blotting | 2 μg / ml | Fig 2 |
| ELISA | 1:500 | Fig 3 |
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C02010034Auto hMeDIP kit x16 (monoclonal mouse antibody)
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C02010031
hMeDIP kit x16 (monoclonal mouse antibody)
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C02010021MagMeDIP Kit
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C02020011
Auto MethylCap kit
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C02020010
MethylCap kit
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C02030030Premium Bisulfite kit
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C15410255-100TET2 Antibody
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C15410311TET3 Antibody
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C15200081-100
5-methylcytosine (5-mC) Antibody - clone 33D3
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C15410204-100
5-Carboxylcytosine (5-caC) Antibody
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C15310200
5-formylcytosine (5-fC) Antibody
Validation Data
Figure 1. hMeDIP results obtained with the monoclonal antibody directed against 5-hmC
hMeDIP (hydroxyMethylated DNA immunoprecipitation) was performed in triplicateon 1 µg fragmented human genomic DNA using 2 µg of the monoclonal antibody against 5-hmC (cat. No. C15200200) and the hMeDIP Kit (cat. No. C02010031). IgG (2 µg) was used a negative IP control. The fragmented DNA was spiked with the internal controls present in the kit (hydroxymethylated DNA (hmeDNA) as a positive and unmethylated DNA (unDNA) as a negative control) prior to performing the IP. QPCR was performed with optimized primer sets, included in the kit, specific for the hydroxymethylated and unmethylated DNA controls. Figure 2 shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. Dot blot analysis using the monoclonal antibody directed against 5-hmC
To demonstrate the specificity of the antibody against 5-hmC (cat. No. C15200200), a Dot blot analysis was performed using an unmodified, a 5-mC and a 5-hmC containing DNA fragment. 100 to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the DNA fragments were spotted on a membrane. The antibody was used at a concentration of 2 µg/ml. Figure 3 shows a high specificity of the antibody for the hydroxymethylated fragment.
Figure 3. Determination of the antibody titer
To determine the titer, an ELISA was performed using a serial dilution of the mouse monoclonal antibody directed against 5-hmC (Cat No. C15200200) in antigen coated wells. The antigen used was the 5-hmC base coupled to KLH. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:40,000.
Target description
5-hydroxymethylcytosine (5-hmC) has been recently discovered in mammalian DNA. This results from the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of oxygenases. So far, the 5-hmC bases have been identified in Purkinje neurons, in granule cells and embryonic stem cells where they are present at high levels (up to 0,6% of total nucleotides in Purkinje cells).
Preliminary results indicate that 5-hmC may have important roles distinct from 5-mC. Although its precise role has still to be shown, early evidence suggests a few putative mechanisms that could have big implications in epigenetics : 5-hydroxymethylcytosine may well represent a new pathway to demethylate DNA involving a repair mechanism converting 5-hmC to cytosine and, as such open up entirely new perspectives in epigenetic studies.
Due to the structural similarity between 5-mC and 5-hmC, these bases are experimentally almost indistinguishable. Recent articles demonstrated that the most common approaches (e.g. enzymatic approaches, bisulfite sequencing) do not account for 5-hmC. The development of the affinity-based technologies appears to be the most powerful way to differentially and specifically enrich 5-mC and 5-hmC sequences. The results shown here illustrate the use of this unique monoclonal antibody against 5-hydroxymethylcytosine that has been fully validated in various technologies.
Documents
| Datasheet 5hmC DATASHEET Monoclonal antibody raised in mouse against 5-hydroxymethylcytosine conjugated to BSA. | Download |
| Exclusive Highly Specific Kits Antibodies for DNA HydroxyMethylation Studies POSTER Cytosine hydroxymethylation was recently discovered as an important epigenetic mechanism. This cy... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
Safety sheets
| 5-hmC antibody mouse SDS GB en | Download |
| 5-hmC antibody mouse SDS US en | Download |
| 5-hmC antibody mouse SDS JP ja | Download |
| 5-hmC antibody mouse SDS DE de | Download |
| 5-hmC antibody mouse SDS BE nl | Download |
| 5-hmC antibody mouse SDS FR fr | Download |
| 5-hmC antibody mouse SDS BE fr | Download |
| 5-hmC antibody mouse SDS ES es | Download |
Publications
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5-Hydroxymethylcytosine in circulating cell-free DNA as a potential diagnostic biomarker for SLE |
RNA m5C oxidation by TET2 regulates chromatin state and leukaemogenesis |
Bioengineering novel AAV9-mGULO-GT for multi-disease gene therapy:Targeting mutated GULO expression to cure scurvy and brain diseases. |
Methylation and expression of glucocorticoid receptor exon-1 variants andFKBP5 in teenage suicide-completers. |
Gene body DNA hydroxymethylation restricts the magnitude oftranscriptional changes during aging. |
The age of bone marrow dictates the clonality of smooth muscle-derivedcells in atherosclerotic plaques. |
Relationship between osteoporosis and osteoarthritis based on DNA methylation |
TET3 prevents terminal differentiation of adult NSCs by a non-catalytic action at Snrpn. |
miR-30a as Potential Therapeutics by Targeting TET1 through Regulation of Drp-1 Promoter Hydroxymethylation in Idiopathic Pulmonary Fibrosis |
Dynamic interplay between locus-specific DNA methylation and hydroxymethylation regulates distinct biological pathways in prostate carcinogenesis |
Hydroxymethylation of microRNA-365-3p Regulates Nociceptive Behaviors via Kcnh2 |
The dual specificity phosphatase 2 gene is hypermethylated in human cancer and regulated by epigenetic mechanisms |
Single-Base Resolution Analysis of 5-Formyl and 5-Carboxyl Cytosine Reveals Promoter DNA Methylation Dynamics. |
Dynamic reprogramming of 5-hydroxymethylcytosine during early porcine embryogenesis. |
Vitamin C induces Tet-dependent DNA demethylation and a blastocyst-like state in ES cells. |
5-Hydroxymethylcytosine is associated with enhancers and gene bodies in human embryonic stem cells. |
5-Hydroxymethylcytosine in the mammalian zygote is linked with epigenetic reprogramming. |