5-Hydroxymethylcytosine (5-hmC) antibody (mAb)

for 5-Hydroxymethylcytosine (5-hmC) antibody (mAb) (Clone 59.1)Background

In addition to this monoclonal, Active Motif offers two polyclonal antibodies that recognize 5-hydroxymethylcytosine, a whole serum version (39769) and a purified IgG version (39791).All are validated for use in methyl DNA immunoprecipitation (MeDIP). For customers that require the ability to quantitate the amount of IgG in the MeDIP reaction, we recommend either this monoclonal or the purified IgG polyclonal (39791). The whole serum version (39769) is very high titre and should be used carefully (0.1 - 0.5 µl per IP) as not to generate high non-specific background. The whole serum version (39769) has been used successfully in immunofluorescence (IF, Ito et al, 2010), and the purified IgG version (39791) is likely to work in this application as well.

DNA methylation is an epigenetic event in which DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide. This modification helps to control gene expression and is also involved in genomic imprinting, while aberrant DNA methylation is often associated with disease. 5-methylcytosine is a modified base that is found in the DNA of plants and vertebrates.
A second type of DNA methylation exists, 5-hydroxymethylcytosine (5-hydroxy methylcytosine, 5-hmC). This results from the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of cytosine oxygenases. This antibody was developed specifically to distinguish 5-hydroxymethylcytosine from 5-methylcytosine as conventional methods (enrichment by antibody or methyl DNA binding protein, enzymatic digestion and bisulfite sequencing) cannot do so.
It is possible that 5-hydroxymethylcytosine (5-hmC) represents a pathway to demethylate DNA, as 5-hydroxymethylcytosine is repaired as mismatched DNA and replaced with unmethylated cytosine.

for 5-Hydroxymethylcytosine (5-hmC) antibody (mAb) (Clone 59.1)Immunogen

This 5-Hydroxymethylcytosine antibody was raised against 5-hydroxymethylcytidine conjugated to KLH and recognizes 5-hydroxymethylcytosine.

for 5-Hydroxymethylcytosine (5-hmC) antibody (mAb) (Clone 59.1)Application Notes

This antibody recognizing 5-hydroxymethylcytosine has been validated for use in dot blot (0.2 µg/ml dilution) and MeDIP (Methyl DNA Immunoprecipitation, 1 - 2 µg per IP).

In MeDIP experiments, the ratio of the % input recovered for the 5-hydroxymethylcytosine sample compared to 5-methylcytosine is 372 : 1. See Figure 2.

for 5-Hydroxymethylcytosine (5-hmC) antibody (mAb) (Clone 59.1)Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 32 mM glycine, 0.07 mM EDTA, 30% glycerol, and 0.035% sodium azide.

for 5-Hydroxymethylcytosine (5-hmC) antibody (mAb) (Clone 59.1)Storage

Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

for 5-Hydroxymethylcytosine (5-hmC) antibody (mAb) (Clone 59.1)Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.