G9a Chemiluminescent Assay Kit
The G9a Chemiluminescent Assay Kit is designed to measure G9a activity for screening and profiling applications. This kit comes in a convenient format, with a plate precoated with histone H3 peptide substrate, recombinant human G9a enzyme (amino acids 785-1210), high specific antibody against the methylated lysine residue of Histone H3, secondary HRP-labeled antibody, S-adenosylmethionine and Methyltransferase Assay Buffer for 100 enzyme reactions.
Figure 1: G9a Chemiluminescent Assay Kit workflow diagram. First, S-adenosylmethionine is incubated with the assay buffer, G9a enzyme, and the inhibitor of choice for one hour in a histone-precoated plate. This is followed by the addition of a primary antibody against Lys(Me). Addition of a chemiluminescent HRP substrate provides a luminescence signal, which directly correlates with the activity of G9a.
Need us to run inhibitor screens or profile your compounds against G9a? Check out our Methyltransferase Screening Services.
This product has been cited 1 time.
- TBST Buffer (1x TBS, containing 0.05% Tween-20), pH 8
- Luminometer or plate reader capable of reading chemiluminescence
- Adjustable micropipettor and sterile tips
- Orbital shaker
Catalog # | Name | Amount | Storage |
51001 | G9a (EHMT2), GST-Tag (Sf9-derived)* | 8 µg | -80°C |
52120 | 20 µM S-adenosylmethionine | 250 µl | -80°C |
52140E | Primary Antibody 5 | 12.5 µl | -80°C |
52130H | Secondary HRP-Labeled Antibody 1 | 10 µl | -80°C |
52160 | 4x HMT Assay Buffer 1 | 3 ml | -20°C |
52100 | Blocking Buffer 4 | 50 ml | +4°C |
0.5 M DTT | 200 µl | -20°C | |
HRP Chemiluminescence Substrate (2 components) | 6 ml each | +4°C | |
96-well module plate (pre-coated with Histone Substrate)** | 1 plate | +4°C |
*The concentration of the protein is lot-specific and will be indicated on the tube.
**Custom coated plates (example, with a lower concentration of Histone Substrate) are available upon request.
G9a, also known as euchromatic histone-lysine N-methyltransferase 2 or EHMT2, belongs to the histone methyltransferase enzyme family. It is involved in mono- and di-methylation of histone H3, at lysine 9 and lysine 27, which control epigenetic regulation of pluripotency during embryogenesis, by depositing H3K9me2 in the promoters of genes like OCT4 (octamer-binding transcription factor 4). It is also involved in neuropathic pain, by regulating a large number of genes in the dorsal root ganglia. G9a has been implicated in cancer cell growth, having a key role in self-renewal of cancer stem cells (CSCs). The presence of CSC populations leads to cancer resurfacing after treatment and they tend to be resistant to typical cancer treatments. The use of inhibitors targeting G9a can result in toxicity, such as in the case of UNC0642 or BIX-01294. The development of newer context-specific inhibitors will provide new therapeutic avenues in cancer therapy.
Dillon S. C., et al., 2005 Genome Biology 6:227.
Haebe J., et al., 2021 Oncogenesis 10: 76.