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Enzymes >
G9a Methyltransferase |
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Prices are in US dollars and valid only for US orders.
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Description:
G9a Methyltransferase methylates lysine 9 (Lys 9) of histone H3 (1-3). Methylation occurs at the e amino group of lysine residues. Methylation of histone H3 Lys 9 is a hallmark of silent chromatin and is globally distributed throughout the heterochromatic regions, such as centromere and telomere (4,5).
Source:
G9a enzyme is expressed from mouse G9A cDNA using a baculovirus expression system (1,2).
Reagents Supplied:
HMT Reaction Buffer (10X)
S-adenosylmethionine (SAM) (32 mM)
Enzyme Properties
Heat Inactivation:
65°C for 20 minutes
Reaction & Storage Conditions
Reaction Conditions:
1X HMT Reaction Buffer
Supplemented with 160 μM S-adenosylmethionine (SAM)
Incubate at
37°C.
1X HMT Reaction Buffer:
50 mM Tris-HCl
5 mM MgCl2
4 mM Dithiothreitol
pH 9.0 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to catalyze the transfer of 1 pmol of methyl group to synthetic peptide substrate representing the first 17 amino acids of histone H3 in a total reaction volume of 25 µl in 10 minutes at 37°C.
Concentration:
2,000 units/ml
Storage Conditions:
50 mM Tris-HCl
100 mM NaCl
5 mM MgCl2
4 mM Dithiothreitol
50% Glycerol
pH 8.0 @ 25°C
Storage Temperature:
-20°C
Notes
General notes:- Storage Note: S-adenosylmethionine (SAM) is stored at -20°C as a 32 mM stolution dissolved in 0.005 M sulfuric acid and 10% ethanol (pH 7.5). Under these conditions, SAM is stable for up to 6 months. SAM is unstable at 37°C and should be replenished in reactions incubated longer than 4 hours. Methylation can be optimized by using fresh SAM.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
Purified free of contaminating proteases.
References
- Tachibana, M. et al. (2001) J. Biol. Chem., 276, 25309-25317.
- Patnaik, D et al. (2004) J. Biol. Chem., 279, 53248-53258.
- Esteve, P.O. et al. (2005) Nucl. Acids Res., 3211-3223.
- Strahl and Allis (2000) Nature, 403, 41-45.
- Noma, K. et al. (2001) Science, 293, 1150-1155.
Reagents Sold Separately
S-adenosylmethionine (SAM)
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