BromoMELT™ Assay Kit

Reaction Biology offers both biochemical and biophysical assays to study epigenetic reader domains. More than 100 assays are available for screening, lead optimization, or selectivity profiling for reader domain inhibitors.

Bromodomains, which belong to the reader category, and recognize acetylated lysine residues on histones and other proteins. Several potent bromodomain compounds have recently been identified, increasing the appreciation of this family’s functional importance and therapeutic potential. At Reaction Biology, we are working towards the complete coverage of the bromodomain family.

  • Extensive coverage of the bromodomain family
  • Test selectivity of any compound within hours in your lab
  • All reagents included
  • All reader domain proteins are produced at our facility and are available for purchase
  • Purchase the BromoMELT™ Assay Kit for in-house screening

Additional Information

Kit description

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.

Workflow:

  1. Prerun (optional)
  2. Prepare PCR master mix, including SYPRO Orange dye.
  3. Pipette reaction plate, including DMSO control, control compound mix, test compound mix in duplicates.
  4. Run qPCR.
  5. Perform data analysis using your instrument’s software.

Download example data.

Download the instruction manual for more detailed information.

Kit Components

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.