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for Recombinant Tet1 protein protein, activeContents
25 µg recombinant Tet1 protein expressed in E. coli supplied in a buffer of 20 mM Tris 8.0, 300 mM NaCl, 2 mM DTT and 5% glycerol.
for Recombinant Tet1 protein protein, activeApplication Notes
Recombinant Tet1 protein is suitable for use in conversion of 5-methylcytosine containing DNA into DNA containing 5-hydroxymethylcytosine. A good starting point is 0.5 to 5 μg of enzyme per assay incubated with 0.5 μg of substrate in 50 mM HEPES, pH 6.8, 50 μM Fe(NH4)2(SO4)2, 2 mM ascorbate, and 1 mM alpha-ketoglutarate for 3 hr at 37°C.
for Recombinant Tet1 protein protein, activeProtein Details
Tet1 is a member of the cytosine oxygenase family of enzymes that convert 5-methylcytosine to 5-hydroxymethylcytosine. Amino acids 1367 to 2039 of mouse Tet1 (accession number ACY38291.1) were expressed in E. coli (MW=95 kDa).
Tet1 activity assay.
Recombinant Tet1 activity measured using an antibody-based activity assay.
Top Panel: 5-hydroxymethylcytosine DNA standards detected with an antibody recognizing 5-hydroxymethylcytosine. Amount of DNA (nanograms) is indicated above each spot.
Bottom Panel: Detection of Tet1 activity. Double-stranded DNA containing 5-methylcytosine was incubated with 5 µg of recombinant Tet1 enzyme (+Tet1), without Tet1 (-Tet1), or unmethylated DNA not incubated with enzyme (DNA) and then spotted and incubated with an antibody recognizing 5-hydroxymethylcytosine to detect conversion of 5-methylcytosine into 5-hydroxymethylcytosine.
Tet1 protein gel.
Tet1 run on an SDS-PAGE gel and stained with Coomassie blue. Top band is Tet1, lower band is a breakdown product.
for Recombinant Tet1 protein protein, activeStorage
Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Recombinant Tet1 protein protein, activeGuarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
