Protein Labeling and Tethering Kit (ybbR)

€1,065

SKU: 00006 Category:

The Protein Labeling and Tethering Kit (ybbR) enables high-resolution single-molecule studies of protein dynamics using optical tweezers. This highly efficient kit supports two main procedures:

  1. Protein labeling with functionalized DNA handles.

  2. Protein tethering to beads for optical tweezers experiments.

The labeling procedure follows a two-step process. First, CoA-modified oligonucleotides are covalently attached to the two ybbR tags fused to the protein of interest using Sfp phosphopantetheinyl transferase. Next, these protein–oligo conjugates are annealed to DNA handles with complementary 5’-overhangs. The kit offers a choice of 517 bp, 1506 bp, or 5039 bp DNA handles (referred to as 0.5 kb, 1.5 kb, and 5 kb, respectively), providing flexibility depending on your experimental needs.

Tethering is achieved using streptavidin- and anti-digoxigenin-coated beads, making the kit fully compatible with dual-trap optical tweezers platforms.

To simplify setup and quality control, the kit includes adenylate kinase (AdK) with two fused ybbR tags, which serves as a reliable control protein for both the labeling and tethering workflows.

Whether you’re new to single-molecule force spectroscopy or looking to streamline your workflow, the Protein Labeling and Tethering Kit (ybbR) minimizes the time, cost, and troubleshooting typically associated with personnel training and protocol optimization.

Need extra material? CoA-oligonucleotides or DNA handles are also available separately for additional labelings or measurements.

The kit enables 2 protein labelings with oligonucleotides (allowing up to 40 experimental sessions on the C-Trap per labeled protein) and supplies DNA handles for 10 experimental sessions on the C-Trap.

Estimated 1-4 weeks delivery time

Specifications

Store at -80°C:

  • CoA Oligonucleotide (36 nt): 2x 20 µl tubes | 100 µM.
    Oligonucleotides for protein labeling using ybbR peptides and Sfp phosphopantetheinyl transferase.

  • Selection of DNA Handles:
    * 0.5 kb (50 ng/µl) | 45 µl.
    * 1.5 kb (150 ng/µl) | 45 µl.
    * 5.0 kb (75 ng/µl) | 65 µl.

  • Sfp Enzyme: 2 x 5 µl tubes | 50 µM.
  • 10x Sfp Reaction Buffer: 50 µl.

  • TCEP (100 mM): 2 x 5 µl tubes.
    Reducing agent.

  • AdK – 6x His (ybbR): 15 µl | 50 µM.
    Adenylate kinease control protein with two ybbR.

Store at 4°C:

Beads are supplied in PBS with 3 mM sodium azide. Exact diameter is indicated on the tube label.

  • Streptavidin-coated polystyrene (⌀ 1.0–1.4 µm): 12 µl | 1.0% (w/v) (typical dilution factor: 1:1000).

  • Anti-digoxigenin-coated polystyrene (⌀ 0.7–0.9 µm): 60 µl | 0.1% (w/v) (typical dilution factor: 1:100).

Product resources

Want to learn more?

Download the protocols:

See our Protein Folding and Conformational Changes application notes.

Want to use this product in practice?

Find out more about the C-Trap here.

Related products