HK-NYS-1 Helix Amine Coupling Kit
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Specifications:
- Product Name: heliX + Y-Structure Kit 1
- Order Number: HK-NYS-1
- Concentration: Refer to Table 1 for specific
concentrations - Buffer: TE40
- For Research Use Only
Product Description:
The heliX + Y-Structure Kit 1 is designed for proximity binding
assays, specifically for Spot 1. It includes new Y-structure
strands and ligands for binary binding in red and green, as well as
cAnchor strand 2 for ternary binding. The kit is intended for
research use only and has a limited shelf life.
Contents and Storage Information:
| Material | Storage Information |
|---|---|
| New Y-structure Red Adapter strand with Ra | – |
| New Y-structure Green Adapter strand with Ga | – |
| New Y-structure Ligand Free Strand (lfs) Red | – |
| New Y-structure Ligand Free Strand (lfs) Green | – |
| cAnchor strand 2 | – |
| Cap Red Green Yellow Yellow Black | – |
| Concentration | Refer to Table 1 for specific concentrations |
| Buffer | TE40 [1] |
Product Usage Instructions:
Preparation:
Step 1:
For higher FRET quality, covalent coupling of ligands is
recommended. Protein can be captured via His-tag using the
HK-NYS-NTA kit.
Step 2:
Mix the Y-structure Adapter strands with Ligand strands in a
specified ratio to build the Y-structure.
Step 3:
Mix the solution from step 2 with cAnchor strand 2 in a
specified ratio.
Step 4:
Follow the example provided for proper vial contents mixing.
Assay Setup in heliOS:
To study ternary complex formation or bispecific small molecules
binding, follow the steps in heliOS:
- Create a New Assay Workflow.
- Add Custom Assay and load Y-Structure FRET Kinetics.
- Modify parameters based on your needs and run the assay.
Suggested assay parameters are available within the heliOS
software.
For binary interaction in red, set LED red to 1 and remember to
set it back to 0 for FRET investigations. Refer to the heliX+ guide
for more details.
To study bispecific antibodies with weak affinities, follow
similar steps in heliOS but choose Kinetics with Functionalization
from the Custom Assay list.
FAQ:
Q: What should I do if I encounter issues with FRET
quality?
A: For improved FRET quality, consider covalent coupling of
ligands or capturing proteins via His-tag using the HK-NYS-NTA
kit.
Q: How should I store the product to ensure its longevity?
A: Store the product as per the storage information provided in
the manual and avoid excessive freeze-thaw cycles by aliquoting the
nanolever.
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heliX +
User Manual
NEW Y-STRUCTURE KIT 1
for proximity binding assay – Spot 1
Dynamic Biosensors GmbH HK-NYS-1 v1.0
Key Features
· Ideal for studying ternary complex formation upon binding of bispecific small molecules (e.g., PROTACs, molecular glues), homo-dimerization and bispecific antibodies with weak affinities.
· The new optimized Y-structure design allows higher FRET sensitivity. · Compatible with heliX® Adapter Chip. · The Y-structure Red Adapter strand carries a moderately hydrophilic red dye (Ra) with a single positive net charge. · The Y-structure Green Adapter strand carries a hydrophilic green dye (Ga) with a single negative net charge. · Green and red dyes can detect binding on each arm via Fluorescence Proximity Sensing (FPS), or report if the
structure is OPEN or CLOSED via sensitive FRET. · Homo-/hetero-proteins can be coupled easily to the arms via exchangeable ligand strands. · The flexible hinge region confines two proteins to a small volume and defined distance.
Product Description
Order Number: HK-NYS-1
Table 1. Contents and Storage Information
Material
New Y-structure Red Adapter strand with Ra
New Y-structure Green Adapter strand with Ga
New Y-structure Ligand Free Strand (lfs) Red for binary binding in green
New Y-structure Ligand Free Strand (lfs) Green for binary binding in red
cAnchor strand 2
Cap Red Green Yellow
Yellow
Black
Concentration 400 nM 400 nM
Amount 5 x 50 µL 5 x 50 µL
500 nM
3 x 100 µL
500 nM
3 x 100 µL
100 nM
5 x 200 µL
For research use only. This product has a limited shelf life, please see expiry date on label. To avoid many freeze thaw cycles please aliquot the nanolever.
Buffer TE40 [1] TE40 [1]
Storage -20°C -20°C
TE40 [1] -20°C
TE40 [1] -20°C
TE40 [1] -20°C
HK-NYS-1 v1.0
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Page 1 of 5
heliX® Adapter Chip Overview
Ternary binding
Binary binding in green
Binary binding in red
HK-NYS-1 v1.0
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Preparation
Step 1
The Ligand strands can be extended at either the 5′ (Ligand strand 1 – Red arm) or 3′ (Ligand strand 2 – Green arm) end with any DNA/RNA sequence. Additionally, they can be crosslinked to a protein of interest through amine coupling using the specialized heliX® Amine Coupling Kits (HK-NYS-NHS-1, which hybridizes to the red arm, and HK-NYS-NHS-2, which hybridizes to the green arm). It is highly recommended to purify the conjugation product with proFIRE® before conducting kinetic studies.
For higher FRET quality, the covalent coupling of the ligands is recommended. TIP Alternatively, the protein can be captured via His-tag using the HK-NYS-NTA kit. For any questions, please
contact the support team at support.dbs@bruker.com.
Step 2
For surface functionalization, the new Y-structure Red Adapter strand harboring the red dye Ra and the new Y-structure Green Adapter strand harboring the green dye Ga need to be pre-hybridized with the Ligand-strands in order to build the Y-structure. In solution hybridization of Y-structure strands:
i. Mix the new Y-structure Red Adapter strand with Ra (400 nM), the new Y-structure Green Adapter strand with Ga (400 nM), conjugated Y-structure Ligand strand 1 (500 nM) and conjugated Y-structure Ligand strand 2 (500 nM) at a 1:1 ratio (v/v).
ii. Incubate the solution of step i) at RT for 2 hours to ensure complete hybridization. Overnight incubation at 4°C is also possible, but it depends on the stability of the conjugated protein.
Step 3
Mix solution of step ii) and cAnchor strand 2 (100 nM) at 1:1 ratio (v/v).
Step 4
Solution is ready to use for heliX® Adapter Chip functionalization.
HK-NYS-1 v1.0
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Page 3 of 5
Example
Required volume for 1 functionalization: 35 µL with a final concentration of 50 nM.
Vial 1
Y-structure Red Adapter strand (400 nM)
4.5 µL
Y-structure Green Adapter strand (400 nM)
4.5 µL
Conjugated Ligand strand 1 (500 nM)
4.5 µL
Conjugated Ligand strand 2 (500 nM)
4.5 µL
Vial 2 cAnchor strand 2 (100 nM)
18 µL
Assay Setup in heliOS
For studying ternary complex formation upon binding of bispecific small molecules (e.g., PROTACs, molecular glues). Go to heliOS > create a New Assay Workflow > add Custom Assay > load Y-Structure FRET Kinetics > modify the parameters based on your needs and run the assay.
Suggested assay parameters (e.g., flow rates, functionalization time, LED power, etc.) are within the heliOS assay.
IMPORTANT
For binary interaction in red, please set LED red 1. However, do not forget to set it back to 0 when FRET interactions are under investigation. For more details, please refer to the heliX+ guide available at this link.
Alternatevely, Y-Structure FRET Kinetics – auto LED assay can be utilized, where the heliX® system automatically adjusts the LED power to optimize the fluorescence signal for a better signal-to-noise ratio. This approach is highly recommended for weak binders or screening applications.
For studying bispecific antibodies with weak affinities (e.g., Hemlibra binding to Factor X and IX) Go to heliOS > create a New Assay Workflow > add simply Kinetics with Functionalization from the Custom Assay list > modify the parameters based on your needs and run the assay.
Antibodies are big proteins which do not allow to bring the two dyes in close proximity, therefore FRET TIP cannot be recorded. This is the reason why classic kinetics workflow and Fluorescence Proximity Sensing
(FPS) is used for detecting binding.
For further questions, please contact the support team at support.dbs@bruker.com.
Useful Order Numbers
Table 2. Order Numbers
Product Name Y-structure Amine coupling kit 1 – Red Y-structure Amine coupling kit 2 – Green New Y-structure Kit 2: for proximity binding assay Spot 2 New Y-structure His Capture Kit
Amount 3 conjugations 3 conjugations 400 nM x 250 µL
500 nM x 200 µL
Order No HK-NYS-NHS-1 HK-NYS-NHS-2 HK-NYS-2
HK-NYS-NTA
HK-NYS-1 v1.0
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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.biosensors@bruker.com Technical Support support.dbs@bruker.com
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.
HK-NYS-1 v1.0
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Documents / Resources
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Dynamic Biosensors HK-NYS-1 Helix Amine Coupling Kit [pdf] Installation Guide HK-NYS-1, HK-NYS-1 Helix Amine Coupling Kit, HK-NYS-1, Helix Amine Coupling Kit, Coupling Kit, Kit |