We designed the BromoMELTTM Assay Kit for profiling of compound binding to a collection of human bromodomain proteins. BromoMELTTM is a thermal stability assay (TSA), where heat-induced protein denaturation exposes hydrophobic surfaces that interact with a dye, thereby increasing its fluorescence. A plot of fluorescence versus the gradually increasing temperature is analyzed to obtain a melting temperature (Tm), represented by an inflection point of the curve. Interaction between the protein and a ligand increases protein stability, leading to an increase in Tm.
- Prerun (optional)
- Prepare PCR master mix, including SYPRO Orange dye.
- Pipette reaction plate, including DMSO control, control compound mix, test compound mix in duplicates.
- Run qPCR.
- Perform data analysis using your instrument’s software.
Download example data.
Download the instruction manual for more detailed information.
Bromodomain Assay Kit Components
- 5x Protein stock plate, 15 uL of each protein
- Reaction buffer, 5x concentrated solution (1 ml)
- 5000x SYPRO Orange (8 uL)
- Control compound mix (5 uL)
- 100% DMSO (10 uL)
- 1x 384 well qPCR plate
- 1x clear seal
- 96 well dispensing guide
- Optional pre-test: 5x pre-test protein, 30 uL
- 1x clear seal, small
Components required but not supplied
- Centrifuge with plate compatible baskets
- 96 well plate
- Real time PCR/qPCR instrument
- Multichannel micropipette
- 400x Stock(s) of Test Compound in DMSO
Purchase the BromoMELT™ assay kit for in-house screening.