DOT1L (KMT4) Chemiluminescent Assay Kit
The DOT1L Chemiluminescent Assay Kit is designed to measure DOT1L activity for screening and profiling applications. The DOT1L Chemiluminescent Assay Kit comes in a convenient format, with a 96-well plate, DOT1L substrate, primary antibody against methylated lysine residue of Histone H3, secondary HRP-labeled antibody, Sadenosylmethionine, methyltransferase assay buffer, and purified DOT1L for 100 enzyme reactions. The key to the DOT1L Chemiluminescent Assay Kit is a highly specific antibody that recognizes methylated K79 residue of H3. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, S-adenosylmethionine is incubated with a sample containing assay buffer and methyltransferase enzyme for two hours. Next, primary antibody is added. Finally, the strips are treated with an HRP-labeled secondary antibody followed by addition of HRP substrate to produce chemiluminescence that can then be measured using a chemiluminescence reader.
Need us to run inhibitor screens or profile your compounds against DOTL (KMT4)? Check out our Methyltransferase Screening Services.
This product has been cited 6 times.
TBST buffer (1x Tris-buffered saline, pH 8.0, containing 0.05% Tween-20)
Luminometer or microplate reader capable of reading chemiluminescence
Rotating or rocker platform
0.5 M DTT
| Catalog # | Component | Amount | Storage | |
| 51005 | DOT1L | 50 µg | -80°C | Avoid freeze/ thaw cycles! |
| 52120 | 100 µM S-adenosylmethionine | 250 µl | -80°C | |
| DOT1L substrate | 250 µl | -80°C | ||
| 52140Y | Primary Antibody 25 | 12.5 µl | -80°C | |
| 52131H | Secondary HRP-labeled Antibody 2 | 10 µl | -80°C | |
| 52180 | 4x HMT Assay Buffer 3* | 3 ml | -20°C | |
| 52100 | Blocking Buffer 4 | 50 ml | +4°C | |
| 79670 | ELISA ECL Substrate A (translucent bottle) |
6 ml | Room Temp. | |
| ELISA ECL substrate B (brown bottle) | 6 ml | Room Temp. | ||
| 79837 | 96-well strip plate | 1 plate | ||
* Add 30 µl of 0.5 M DTT before use.
1. Min, J., et al. Cell 2003; 112:711-723.
2. Daigle, S.R., et al. Cancer Cell 2011, 20:53-65.
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