?-Glucosyltransferase enzyme

by Active Motif
Available SKUs: 55012 |
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Description

for Recombinant B-Glucosyltransferase enzymeContents

200,000 Units ?-Glucosyltransferase enzyme
10X ?-Glucosyltransferase Reaction Buffer
50 mM UDP-Glucose
1 M DTT.

for Recombinant B-Glucosyltransferase enzymeApplication Notes

The ?-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

?-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.

 
?-Glucosyltransferase protection of 5-hmC DNA from GlaI digestion

?-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 ?-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 guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

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