ICH ELECT Services – Determine Cellular Potency Against Novel Targets
Unlike other biochemical or cell-based epigenetic assays that are limited by the availability of specific antibodies or reagents that detect histone modifications, InCELL Hunter bromodomain and methyltransferase assays are non-antibody-based, high-throughput friendly chemiluminescent assays and can provide a direct measure of compound permeability and binding to a proteins’ catalytic domain in live cells.
Assay Performance
BRD4(1)
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G9a Methyltransferase
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InCELL Hunter BRD4(1) cells (Left panel) and G9a Methyltransferase cells (Right panel) stimulated with the indicated compounds for 6 hours. Signal was detected using the InCELL Hunter Detection kit. The data demonstrates that an increase in signal is specific to a molecule that binds the target.
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Plate Uniformity
| C0638 |
Hill Slop |
IC50 (nM) |
S:B |
| CRC1 |
1.391 |
66 |
13.6 |
| CRC2 |
1.501 |
80 |
12.5 |
| CRC3 |
1.384 |
98 |
13.1 |
| CRC4 |
1.381 |
94 |
13.1 |
| CRC5 |
1.354 |
90 |
11.2 |
| CRC6 |
1.247 |
106 |
12.1 |
| CRC7 |
1.407 |
102 |
12.0 |
| CRC8 |
1.393 |
76 |
12.0 |
InCELL Hunter G9a Methyltransferase cells were plated in 384-well format and 8 replicates of a 10-point control response curves (run in duplicate ) were performed. Assay performance was consistent across the entire plate
ICH ELECT Panel
Listed below are the assays currently available for screening and profiling.
Bromodomain Assays
| Assay Name ▲ | Common Name | Accession Number |
| BAZ2A | | NM_013449.3 |
| Brd2(1) | BRD2 | NM_005104.3 |
| BRD2(1,2) | BRD2 | NM_005104.3 |
| BRD3(1) | BRD3 | NM_007371.3 |
| BRD3(1,2) | BRD3 | NM_007371.3 |
| Brd4(1) | BRD4 | NM_058243 |
| BrdT(1) | BRDT | NM_001242806 |
| BRDT(1,2) | BRDT | NM_001726 |
| EP300 | | NM_001429.3 |
| SMARCA4 | | NM_001128844 |
| TAF1L(2) | | NM_153809.2 |
Histone Methyltransferase Assays
Data Analysis & Interpretation
Definitions
Percent Binding
The results for single concentration (primary screen) for tested compound(s) are reported in your study report and spreadsheets as '% Binding' and are calculated in the following manner:
% Binding =
100% x
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Mean RLU (test sample)- Mean RLU(vehicle control)
Mean MAX RLU (control ligand)- Mean RLU (vehicle control)
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test sample = client supplied compound
vehicle control = DMSO (0% activity)
control ligand = control compound (100% activity)
Dose Response Curve (EC50)
The results for 10-point dose response curve compound/target interactions are reported in the study report and spreadsheets as cellular EC50s, which are values derived using the following equation:
