Invitrogen™ BacMam Histone H3K4me3 Cellular Assay Kit

The BacMam Histone H3K4me3 Cellular Assay is a high-throughput screening (HTS) compatible TR-FRET assay for the interrogation of trimethylation at lysine 4 on histone H3 in a cellular format. Baculovirus-mediated gene delivery (BacMam) provides a convenient tool for the expression of GFP-Histone H3 fusion protein in your cell background of interest. Coupled with a terbium (Tb)-labeled site-specific antibody, the resulting cellular assay can be used to identify inhibitors of methyltransferases and demethylases acting on lysine 4 of histone H3.

The kit includes:

• BacMam Histone H3 Reagent
• LanthaScreen™ Tb-anti-histone H3K4me3 Antibody
• 6X LanthaScreen™ Cellular Lysis Buffer
• Instrument Control Terbium TR-FRET kit

The BacMam Histone H3K4me3 Cellular Assay Kit Lets You:

• Use your cell background of choice with the portability of BacMam
• Identify more relevant inhibitors since the methyl transferases and demethylases are in their natural protein complexes
• Conserve precious cell samples with the miniaturizable homogenous assay format
• Improve data quality with the advantages of TR-FRET

Get More Physiologically Relevant Results

The BacMam Histone H3K4me3 Cellular Assay allows the investigation of trimethylation at lysine 4 on histone H3 with your choice of cellular backgrounds including primary cells. This enables screening for potential inhibitors of methyl transferases and demethylases associated with Lys4 trimethylation in their natural complexes in a physiologically relevant cell type. Please note that the LanthaScreen™ Tb-anti-Histone H3K4me3 antibody has been shown to have higher affinity for trimethylation compared to dimethylation at lysine 4 using a histone H3 peptide; however, when interpreting your results, it is important to consider that the antibody does show some affinity for the dimethylated peptide. Further discrimination between di- and trimethylation can be accomplished by using our complimentary BacMam Histone H3K4me2 Cellular Assay Kit (Cat. No. A14164), which is specific for dimethylation.

BacMam Convenience Saves Samples and Time

Assays are run in a fully homogenous, addition-only format without any of the washing, lysate transfer, or separation procedures required for traditional methods such as Western blotting and ELISA, making them ideal for HTS applications. By using the same BacMam reagent for GFP-Histone H3, other histone H3 modifications (Ph, Me, Ac) can be monitored just be exchanging the Terbium (Tb) labeled antibody. In addition, the application of the LanthaScreen™ technology includes all the advantages of TR-FRET detection including reduced data noise, less interference from fluorescent compounds, and high sensitivity, allowing the use of fewer cells than traditional Western or ELISA methods.