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Cat. No.: RTET2-10 (for 10μg)

Cat. No.: RTET2-50 (for 50μg)

Description

Recombinant Ten-Eleven Translocation 2 (TET2), also known as Tet Methylcytosine Dioxygenase 2 or TET Methylcytosine Dioxygenase 2, is an Fe²⁺ and α-ketoglutarate (α-KG)-dependent methylcytosine dioxygenase. TET2 catalyzes a series of oxidation reactions that convert 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine (5-hmC), and subsequently into 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC).

TET2 is applicable in next-generation sequencing (NGS)-based methylation analysis, including enzymatic methyl sequencing (EM-seq), Tet-assisted bisulfite sequencing (TAB-seq), and TET-assisted pyridine borane sequencing (TAPS) [2].

Whole Genome Bisulfite Sequencing (WGBS) has long been considered the gold standard for mapping DNA methylation. However, the harsh chemical reactions involved in bisulfite treatment can damage DNA, leading to fragmentation, loss, and pronounced GC bias in methylated regions. Enzyme-based methylation sequencing methods overcome these limitations.

EM-seq constructs whole-genome methylation libraries through two enzymatic steps:

1. Conversion Step: Recombinant TET2 and T4-BGT (T4-phage beta-glucosyltransferase) are used to convert 5-mC and 5-hmC into 5-caC and β-glucosyl-5-hydroxymethylcytosine (5-gmC), respectively. These modified cytosines (5-caC and 5-gmC) are resistant to deamination by APOBEC3A.

2. Deamination Step: The deaminase APOBEC3A selectively deaminates unmodified cytosines (C) into uracils (U), without affecting 5-caC and 5-gmC. This enables clear differentiation between modified and unmodified cytosines.

Thus, the combined use of TET2, T4-BGT, and APOBEC3A enables specific detection of 5-mC and 5-hmC.

 Applications

Recombinant TET2 is suitable for specific detection of 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) in various next-generation sequencing (NGS) methods, including:

• Enzymatic Methyl Sequencing (EM-seq)
• Tet-Assisted Bisulfite Sequencing (TAB-seq)
• TET-Assisted Pyridine Borane Sequencing (TAPS)

It is also applicable in:

• Enzyme kinetics studies
• Inhibitor screening
• Selectivity analysis

 Source

Expressed in Escherichia coli using a human TET2 gene construct and purified to high quality.

Purity

• Free of DNase (endonuclease and exonuclease activity)
• Free of RNase
• Free of phosphatase
• Purity >90%  

Storage 

Store at –20ºC. Stable for at least two years under recommended conditions.

Precautions 

• Keep the product on ice or in an ice bath during use. Return to –20ºC storage immediately after use.
• This product is intended for use by qualified professionals in scientific research only. It is not for use in clinical diagnosis or treatment, food or pharmaceutical applications, and must not be stored in residential settings.
• For your safety and health, please wear a lab coat and disposable gloves when handling this product.

Related:

• Recombinant APOBEC3A (A3A)
• Recombinant Ten-Eleven Translocation 2 (TET2)
• T4 Phage β-Glucosyltransferase (T4-BGT)

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Only for research and not intended for treatment of humans or animals

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