5-Hydroxymethylcytosine (5-hmC) antibody (rAb) (Clone RM236)

Host / Isotype

Rabbit / IgG

Reactivity

Human, Not Species Specific

Applications

DB, IF, IHC

CloneNo.

RM236

Cat No : 91309

Synonyms

-Hydroxymethylcytidine, 5-hydroxymethylcytosine, 5-hmC, 5-Hydroxy methylcytidine, 5-hydroxy methyl cytidine, 5-hydroxymethylcytosine, 5-hmc, 5 hmc, 5hmc, hydroxy methyl cytosine, RM236

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Validation Data Gallery

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  Dot blot of 5-Hydroxymethylcytosine (5-hmC) antibody (rAb).Membrane was pre-spotted with 50, 5, and 0.5 ng/dot of double stranded 5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine (5-mC) DNA, and unmethylated DNA. The pre-spotted membrane was then blotted with antibody at 0.2 ug/ml.

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  MeDIP of 5-Hydroxymethylcytosine (5-hmC) antibody (rAb).MeDIP was performed using 5-Hydroxymethylcytosine antibody (Clone RM236) at a 10:1 DNA:Ab ratio. 1 ng of unmethylated, 5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine (5-hmC) DNA standard (897 bp) was spiked in 1 ug of genomic DNA isolated from HeLa cells as the control. Realtime PCR was then performed to determine the capture of DNA standard as in % of input.

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  Immunofluoresence staining of HeLa cells using 5-Hydroxymethylcytosine antibody (Clone RM236) at 0.5ug/mL (red). Actin filaments are labeled with fluorescein phalloidin (green). HeLa cells were fixed with 4% paraformaldehyde and permeabilized with methanol (−20°C) before treatment with 2 N HCl for 30 min at 37 °C to denature DNA.

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Product Information

• Tested Applications: DB, IF, IHC

  Validated Applications: DB: 0.2 - 1 ug/ml IHC: 0.1 - 1 ug/ml ICC: 0.5 - 2 ug/ml MeDIP: 0.2 - 2 ug/ml

• Tested Reactivity: Human, Not Species Specific
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: BSA-conjugated 5-hydroxymethylcytosine.
• Full Name: 5-Hydroxymethylcytosine (5-hmC) antibody (rAb) (Clone RM236)
• Synonyms: -Hydroxymethylcytidine, 5-hydroxymethylcytosine, 5-hmC, 5-Hydroxy methylcytidine, 5-hydroxy methyl cytidine, 5-hydroxymethylcytosine, 5-hmc, 5 hmc, 5hmc, hydroxy methyl cytosine, RM236
• GenBank accession number: N/A
• Purification Method: Protein A Chromatography
• Buffer: PBS with 50% glycerol, 1% BSA, and 0.09% sodium azide. Sodium azide is highly toxic.
• Storage: Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Background Information

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.