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| |
Mismatch Repair Enzymes
E. coli MutY Enzyme
Mismatch Repair
Oligonucelotides
| E. coli Mut Y acts together
with Fpg to prevent the potentially mutagenic consequences of 8-oxo-dG
lesions. The 8-oxo-dG lesions that escape repair by Fpg frequently pair
with A during DNA replication, producing an 8-oxo-dG:A mispair. Mut Y
removes the A from this base pair to initiate base excision repair. In
the absence of Mut Y, DNA replication past the 8-oxo-dG:A mismatch
results in thymine incorporation opposite the adenine in one of the
daughter strands, creating a fixed mutation. Mut Y has an associated AP
lyase activity. |
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|
|
|
| Purified from E. coli
containing a recombinant plasmid harboring the E. coli
MutY gene. |
| |
| One unit is the amount of enzyme
required to cleave 1 pmole of an oligonucleotide duplex
containing an A/G mismatch in 1 hour at 37°C. Only the strand
with the A is cleaved. |
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| 1X REC™
Buffer 9 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 1 mM EDTA, 1 mM
EGTA, and 0.1 mM DTT), 4 pmoles 32P hypoxanthine
oligonucleotide, 4 pmoles complementary oligonucleotide, and
serial dilutions of enzyme in a reaction volume of 20 µl.
Incubate for 1 hour at 37°C. Reaction products are resolved by
20% denaturing PAGE. |
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| • |
MIDAS™
 |
| • |
MIDAScan™
|
| • |
Mismatch
cleavage assays |
|
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| Cat# |
Componenet |
Size |
4000-500-01
|
Mut Y
|
500 Units
|
| 3900-500-04
|
10X REC™
Buffer 4 |
1 ml
|
| 4019-1
|
REC™
Water |
1 ml
|
| 4017-500
|
3X Alkaline
Loading Buffer |
500 µl
|
| 3800-100-01
|
MUA
Oligonucleotide |
100 pmol
|
| 3800-100-05
|
MLA
Oligonucleotide |
100 pmol
|
| 3800-100-06
|
MLT
Oligonucleotide |
100 pmol
|
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| Freeze in working aliquots at -20°C
in a manual defrost freezer to avoid repeated freeze-thawing.
Enzyme is supplied in 10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 1 mM
EDTA, 0.1 mg/ml BSA, and 50%(v/v) glycerol. |
| |
| MutY DNA glycosylase recognizes A/G
and A/8-oxo-dG mismatches in duplex DNA and cleaves the strand
containing the A. The opposite strand is not cleaved. |
|
Products
|
4000-500-EB |
E. coli
MutY Enzyme, 500 Units and 10X REC Buffer 4, 1 ml |
|
4000-500-K |
E. coli MutY Kit, 500 Units |
Products
|
3800-100-A |
A
Mismatch Set (8-oligonucleotides), 100 pmol each |
|
3800-100-T |
T
Mismatch Set (8-oligonucleotides), 100 pmol each |
|
3800-100-G |
G
Mismatch Set (8-oligonucleotides), 100 pmol each |
|
3800-100-C |
C
Mismatch Set (8-oligonucleotides), 100 pmol each |
|
3810-100-A |
A
Mismatch Set (8-oligonucleotides), 100 pmol each |
|
3810-100-T |
T
Mismatch Set (8-oligonucleotides), 100 pmol each |
|
3810-100-G |
G
Mismatch Set (8-oligonucleotides), 100 pmol each |
|
3810-100-C |
C Mismatch Set (8-oligonucleotides), 100 pmol each |
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+32 (0) 16
58 90 45 
+32 (0) 16
50 90 45
