Biomolecular interactions

Specific interactions between biomolecules are studied by using highly charged 96-well plates.  For instance, negatively charged DNA, RNA or proteins (the first biomolecule) are bound to the highly positively charged plates.   After the plates are blocked with the Nucleic Acid Binding Blocker or BSA, a second biomolecule is added. The second biomolecule will also bind to the plate if it interacts with the first biomolecule. The second molecule can be detected by enzymatic assays if it is an enzyme, fluorescence assay if it is DNA or RNA, or ELISA if its antibody is available.  If the second molecule is fluorescence or isotope labeled, the labeled signals can be detected. We provide highly charged 96-well black or transparent polystyrene plates for various applications.

Examples:

DNA-binding plates: DNA1 (a 40-mer DNA oligo) and DNA2 (a 60-mer DNA oligo) form double strand DNA.  After DNA1 binding to the plate, the plate is treated with the DNA-binding Blocker (Catalog number NBP96N), then DNA2 is added to interact with the bound DNA1.  The double strand DNA is detected by PicoGreen fluorescence dye. Four parallel data sets are plotted in the figure.  

Protein-binding plates: Protein1 (primase) specifically interacts with Protein 2 (helicase).  After Protein 1 binding to the plate, the plate is treated with Blocker (1% BSA), then Protein2 is added to interact with the bound Protein 1.  Protein 2 is measured by its ATPase activity in a time-course experiment.

 

DNA-binding plates

         96-well DNA–binding Plates (black)

         96-well DNA–binding Plates (transparent)
         Nucleic Acid Binding Blocker

Protein-binding plates

         96-well Anionic Protein-binding Plates (black)

         96-well Anionic Protein-binding Plates (transparent)

         96-well Cationic Protein-binding Plates (black)

         96-well Cationic Protein-binding Plates (transparent)   

 

References:

  1. Arora S et al, Downregulation of XPF–ERCC1 enhances cisplatin efficacy in cancer cells, DNA Repair, Volume 9, Issue 7, Pages 745-753 (2010).
  2. Arora S, Identifying and Targeting Cellular Mechanisms to Enhance Cisplatin Chemotherapeutic Response in Cancer.  PhD dissertation, The University of Toledo (2012).
  3. Sawanta A et al, Role of mismatch repair proteins in the processing of cisplatininterstrand cross-links. DNA Repair 35: 126–136 (2015).