This kit uses cysteine-maleimide chemistry to attach DNA handles to proteins of interest and perform optical tweezers measurements of protein folding properties and protein conformational changes.
In a first step, short maleimide-modified oligonucleotides (36 bp) react with specifically incorporated cysteines of the protein. In a second step, the protein-attached oligonucleotides anneal with complementary 5’-overhangs of two 520 bp DNA handles, to form a protein–handle complex that is ready for tethering. The kit includes purified AdK with specifically incorporated cysteines that can be used as positive control and quality assurance for each step of the labeling procedure.
A schematic illustration representing three-domain protein tethered to two optically trapped beads through DNA handles. Extending or retracting protein structure through optical manipulation causes the protein to unfold and fold, respectively. Read more here: : https://lumicks.com/application/protein-folding-optical-tweezers-fluorescence-microscopy/
- Maleimide-modified oligos (36 bp)
- Biotin- and digoxigenin-modified DNA handles (520 bp per handle)
- Reducing agent TCEP
- 6xHis tagged AdK protein
- Buffer 40 mM Tris HCl, 50 mM NaCl
- Streptavidin coated polystyrene beads (1.76 µm)
- Anti-digoxigenin coated polystyrene beads (0.75 µm)
- Estimated for 2 weeks of measurements (handles and beads, more can be ordered as Protein handles kit (recurring))
- Estimated for 8 weeks of measurements (labeled protein and control)
- Estimated 1-3 weeks delivery time