helix+ User Manual
HIS CAPTURE KIT 2
for ligand strand 2 with green dye Ga - only for Y-structure
Dynamic Biosensors GmbH | Order Number: HK-NTA-2 v1.0
Key Features
- This kit is designed for capture of histidine-tagged proteins (His6 or His10) using Tris-NTA.
- Includes Ligand strand 2 with Tris-NTA for 20 regenerations.
- For functionalization of ligand strand 2 (green arm) of the Y-structure.
- Used in combination with HK-NTA-1 for the red arm of the Y-structure, it is ideal for dimerization projects.
- Homo-/hetero-proteins can be coupled easily to the arms via his-tag capture.
- Compatible with heliX® Adapter Chip, both Spot 1 and Spot 2
heliX® Adapter Chip Overview
The heliX® Adapter Chip features 2 spots with 2 different anchor sequences for DNA-encoded addressing. Spot 1 is functionalized with the capture molecule while Spot 2 is used as real-time reference.
Single Capture
The 'Single Capture' diagram illustrates the functionalization of a single spot on the heliX® Adapter Chip. Spot 1 is shown with 'Anchor strand 1' and is functionalized with 'Ligand strand 2 (L2) - NTA', indicated by a green dye. A captured histidine-tagged protein is shown bound to the NTA ligand. Spot 2, equipped with 'c-Anchor strand 2', is designated as a real-time reference. The diagram indicates the 'chip surface'.
Double Capture
The 'Double Capture' diagram depicts a Y-structure setup on the heliX® Adapter Chip. Spot 1 features 'Anchor strand 1' and 'c-Anchor strand 2', functionalized with 'Ligand strand - NTA' (red dye) on one arm and 'Ligand strand 2 (L2) - NTA' (green dye) on the other. Two histidine-tagged proteins are shown captured, one on each arm of the Y-structure. Spot 2, with 'c-Anchor strand 2', serves as a reference. The diagram indicates the 'chip surface'.
Product Description
Order Number: HK-NTA-2
Contents and Storage Information
| Material | Cap | Concentration | Amount | Buffer | Storage |
|---|---|---|---|---|---|
| NTA - Ligand strand 2 | Purple | 500 nM | 2 x 100 μL | TE40 [1] | -20°C |
| Loading Solution (NiCl2) | Transparent | 10 mM | 5 x 1500 μL | TE40 [1] | -20°C |
| Imidazole Solution | Transparent | 250 mM | 10 x 2000 μL | TE140 [2] | -20°C |
For research use only.
This product has a limited shelf life; please see expiry date on label. After preparation of ready-to-use solution, the expiry date is 6 months.
Preparation
Step 1: Hybridization of Y-structure strands
For surface functionalization, the Y-structure Red Adapter strand (harboring the red dye Ra) and the Y-structure Green Adapter strand (harboring the green dye Ga) need to be pre-hybridized with the NTA - Ligand strand and NTA - Ligand strand 2 to build the Y-structure. In solution hybridization:
- Mix Y-structure Red Adapter strand with Ra (400 nM), Y-structure Green Adapter strand with Ga (400 nM), NTA Ligand strand (500 nM), and NTA - Ligand strand 2 (500 nM) at a 1:1 ratio (v/v).
- Incubate the solution from step i) at room temperature (RT) for at least 2 hours to ensure complete hybridization. Overnight incubation at 4°C is also possible.
Step 2: Mix with cAnchor strand 2
Mix the solution from step ii) with cAnchor strand 2 (100 nM) at a 1:1 ratio (v/v).
Step 3: Ready for functionalization
The solution is now ready to use for heliX® Adapter Chip functionalization.
Example
Required volume for one functionalization for double capture on Y-structure (Spot 1): 35 μL with a final concentration of 50 nM.
| Red Adapter strand with Ra (400 nM) | Green Adapter strand with Ga (400 nM) | NTA - Ligand strand (500 nM) | NTA - Ligand strand 2 (500 nM) | cAnchor strand 2 (100 nM) |
| 4.5 μL | 4.5 μL | 4.5 μL | 4.5 μL | 18 μL |
Assay Setup in heliOS
This specific kit requires a customized method for His-tag capture plus Y-Structure FRET Kinetics. This method is not currently provided among the verified assays. An advanced heliOS user can create it by applying default parameters from existing workflows. For assistance in creating the new method, please contact the support team at support@dynamic-biosensors.com.
TIP: As the stability of his capture is affected by the protein, in case of long dissociations, consider using the classic conjugation approach.
Useful Order Numbers
| Product Name | Comment | Order No |
|---|---|---|
| heliX® Adapter Chip | Chip with 2 detection spots | ADP-48-2-0 |
| Y-Structure Kit 1: for proximity binding assay on Spot 1 | For studying ternary complex formation | HK-YS-1 |
| Y-Structure Kit 2: for proximity binding assay on Spot 2 | For studying ternary complex formation | HK-YS-2 |
| His Capture Kit: with red dye (Ra) | For capturing of histidine-tagged proteins using Tris-NTA on the red arm of the Y-structure. | HK-NTA-1 |
Contact
Dynamic Biosensors GmbH
Perchtinger Str. 8/10
81379 Munich
Germany
Bruker Scientific LLC
40 Manning Road, Manning Park
Billerica, MA 01821
USA
Order Information: order@dynamic-biosensors.com
Technical Support: support@dynamic-biosensors.com
Website: www.dynamic-biosensors.com
Instruments and chips are engineered and manufactured in Germany.
©2025 Dynamic Biosensors GmbH
For Research Use Only. Not for use in clinical diagnostic procedures.
[1] TE40: 10 mM Tris, 40 mM NaCl, 0.05 % Tween20, 50 μM EDTA, 50 μM EGTA
[2] TE140: 10 mM Tris, 40 mM NaCl, 0.05 % Tween20, 50 μM EDTA, 50 μM EGTA
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