5-Hydroxymethylcytosine (5-hmC) Monoclonal Antibody [HMC/633]

Background
5-hydroxymethylcytosine (5-hmC), as a sixth DNA base with functions in transcription regulation, has been detected to be abundant in human and mouse brain and embryonic stem (ES) cells. In mammals, it can be generated by oxidation of 5-methylcytosine (5-mC), a reaction mediated by the ten-eleven translocation (TET) family of 5-mC hydroxylases. 5-hmC was demonstrated to play an important and different role than 5-mC in the regulation of DNA methylation, chromatin remodeling, and gene expression.

Concentration
1 mg/ml

Description
Rat monoclonal antibody to 5-Hydroxymethylcytosine (5-hmC), clone HMC/633

Purification
Protein A Purified

Specificity
Modified base 5-hydroxymethylcytosine (5-hmC), a broad range of species

Isotype
IgG2a

Formulation
10 mM phosphate buffer, 150 mM NaCl, pH 7.4, 0. 05% sodium azide

Storage
4°C. For long-term storage, aliquot and store at -20°C. Avoid repeated freezing and thawing. Multiple freeze/thaw cycles may result in decreased performance. Stable for 6 months from the date of shipment

Handling Recommendations
For maximum recovery of the products, centrifuge the vial prior to opening the cap

Alternative Names
5-hydroxymethylcytidine, anti-5-hydroxymethylcytidine, anti-5-hydroxymethylcytosine, 5hmC, 5hmeC, 5-hmeC 

Application
Immunofluorescence: 1:100 – 1: 500
Immunohistochemistry: 1:100 – 1: 500
ELISA: 1:500 – 1:1000
Immunoprecipitation: 1:100 – 1: 200

Fig. 1. Dot blot assay shows that 5-Hydroxymethylcytosine Antibody specifically recognized 5-hydroxymethyl Cytidine (hmC). Lane 1: 10 pmol, Lane 2: 2 pmol; Lane 3: 0.4 pmol.

Fig. 2. Selective enrichment of hydroxymethyated DNA with an anti-5-hydroxymethylcytidine antibody (Cat. No. A-1019). 50 pg of unmethylated, methylated, and hydroxymethylated DNA control were each spiked into fragmented human genomic DNA (500 ng). hMeDIP was processed with the 5-hmC antibody. Eluted DNA was analyzed by real time PCR to detect the presence of spiked control DNA. %input represents the amount of recovered control DNA and was calculated based on the Cts.