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for Recombinant B-Glucosyltransferase enzymeContents
500 Units β-Glucosyltransferase enzyme
10X β-Glucosyltransferase Reaction Buffer
50 mM UDP-Glucose
1 M DTT
for Recombinant B-Glucosyltransferase enzymeBackground
In mammalian genomes, DNA methylation usually occurs at the fifth carbon of cytosine residues. The recent discovery that the TET family of iron-dependent deoxygenases can convert 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine (5-hmC) has raised questions about the functional relevance of 5-hmC in mammalian genomes. The β-Glucosyltransferase enzyme serves as a valuable tool to analyze 5-hmC by enabling modification of the 5-hydroxymethylcytosine residue with the addition of a glucose moiety. This modification can help with the differentiation of 5-mC from 5-hmC.
for Recombinant B-Glucosyltransferase enzymeApplication Notes
The β-Glucosyltransferase enzyme can be used to glucosylate 5-hmC DNA. Glucosylated DNA can be used as a differentiator with glucosyl-sensitive restriction enzymes. The use of a labeled [14C] UDP-Glucose donor would enable labeling of the 5-hmC residues.
for Recombinant B-Glucosyltransferase enzymeProtein Details
β-Glucosyltransferase will transfer the glucose moiety from the UDP-Glucose (uridine diphosphoglucose) to 5-hydroxymethylcytosine residues in double-stranded DNA.
One unit is defined as the amount of enzyme required to fully protect 0.5 µg of 5-hmC DNA Standard (Catalog No. 55008) in 1 hour at 37°C in a total reaction volume of 50 µl from cleavage by 4 units of GlaI restriction enzyme in an incubation of 16 hours at 30°C. A pilot titration of enzyme is recommended for protection of genomic DNA.
β-Glucosyltransferase enzyme protection of 5-hmC DNA shown by agarose gel.
0.5 µg of a 5-hmC Methylated DNA Standard (Catalog No. 55008) was incubated in the absence or presence of 1 unit β-Glucosyltransferase enzyme for 1 hour at 37°C. Following column purification, reactions were treated with 4 units of glucosyl-sensitive restriction enzyme GlaI and run on a 2.5% agarose gel.
for Recombinant B-Glucosyltransferase enzymeStorage
Lyophilized proteins can be stored at -20°C or -80°C, preferably desiccated. 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 B-Glucosyltransferase enzymeGuarantee
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.
