dynamic BIOSENSORS DS-6 Dye Scouting Kit User Manual
- September 6, 2024
- dynamic BIOSENSORS
Table of Contents
dynamic BIOSENSORS DS-6 Dye Scouting Kit
Key Features
- Adapter strand 1 – Ra and Adapter strand 2 – Ra – lfs (ligand free strand) for functionalization of heliX
Adapter
Chip on Spot 1 and Spot 2, respectively.
- Compatible with heliX
Adapter Chip.
- Includes Adapter strands for 10 regenerations for each dye.
- Ideal for MIX&RUN sample preparation.
- This kit contains Adapter strand 1 with 6 different dyes and Adapter strand 2 with 6 different dyes already prehybridized with ligand free strand.
- Dye Scouting enables to screen for the most sensitive fluorophore for your application.
Product Description
Order Number: DS-6
Table 1. Contents and Storage Information
| Material | Cap | Concentration | Amount | Buffer | Storage |
|---|---|---|---|---|---|
| Adapter strand 1 – Ra | Black | 400 nM | 100 µL | TE40 [ 1] | -20°C |
| Adapter strand 1 – Rb | Black | 400 nM | 100 µL | TE40 [ 1] | -20°C |
| Adapter strand 1 – Rc | Black | 400 nM | 100 µL | TE40 [ 1] | -20°C |
| Adapter strand 1 – Ga | Black | 400 nM | 100 µL | TE40 [ 1] | -20°C |
| Adapter strand 1 – Gb | Black | 400 nM | 100 µL | TE40 [ 1] | -20°C |
| Adapter strand 1 – Gc | Black | 400 nM | 100 µL | TE40 [ 1] | -20°C |
| Adapter strand 2 – Ra – lfs | White | 200 nM | 100 µL | TE40 [ 1] |
-20°C
Adapter strand 2 – Rb – lfs| White| 200 nM| 100 µL| TE40 [ 1]|
-20°C
Adapter strand 2 – Rc – lfs| White| 200 nM| 100 µL| TE40 [ 1]|
-20°C
Adapter strand 2 – Ga – lfs| White| 200 nM| 100 µL| TE40 [ 1]|
-20°C
Adapter strand 2 – Gb – lfs| White| 200 nM| 100 µL| TE40 [ 1]|
-20°C
Adapter strand 2 – Gc – lfs| White| 200 nM| 100 µL| TE40 [ 1]|
-20°C
- For research use only.
- This product has a limited shelf life, please see the expiry date on the label.
- To avoid many freeze-thaw cycles please aliquot the nanolever.
heliX Adapter Chip Overview
- 2 spots with 2 different anchor sequences for DNA-encoded addressing.
Preparation | MIX&RUN
In-solution hybridization of adapter and ligand strands:
- Mix Adapter strand 1 – dye (400 nM) and conjugated Ligand strand of interest (500 nM) at a 1:1 ratio (v/v).
- Incubate the solution of step 1 at RT at 600 rpm for 30 min to ensure complete hybridization.
- Mix the solution of step 2 and Adapter strand 2 – dye – lfs (200/250 nM) at a 1:1 ratio(v/v).
- The solution is ready to use for biochip functionalization.
- The stability of the solution is related to the stability of the ligand molecule.
Example
Required volume for 3 functionalizations: 40 µL with a final concentration of
100 nM.
| Vial 1 | Vial 2 |
|---|
Adapter strand 1 – dye
(400 nM)
| Conjugated Ligand strand
(500 nM)
| Adapter strand 2 – dye – lfs
(200/250 nM)
10 µL| 10 µL| 50 µL
- After incubation time, mix vial 1 and vial 2 to obtain 40 µL of ready-to-use DNA solution.
Dye Scouting Information
The two standard fluorescent probes for switchSENSE® measurements are the red and green dye Ra and Ga, respectively. Fluorescence proximity sensing is based on the influence of changes in the local environment on the fluorescence signal of the dye in close proximity. This effect depends on the chemical nature of the dye and the interaction partner. Therefore, a different dye may yield a higher signal response depending on the type of interaction. This dye scouting kit enables to screen for the most sensitive fluorophore for your application. Three red and three green fluorophores with different chemical properties are available to choose from for your switchSENSE® assay. The basic differences of the dyes in terms of net charge and hydrophobicity are depicted in Figure 1.
Figure 1. Overview of the switchSENSE®
dye properties. Six fluorophores – three red dyes (Ra, Rb, Rc) and three green
dyes (Ga, Gb, Gc) – are available. Depending on the interaction partner,
different fluorescent probes may obtain different signal responses. Dye
scouting allows to quickly screen for maximum signal amplitudes.
Assay Setup in heliOS
Go to heliOS > create a New Assay Workflow > add Custom Assay > load Dye Scouting > modify the parameters based on your needs and run the assay. Suggested assay parameters (e.g., flow rate, time, LED power, etc.) are within the heliOS assay. As default setting all six fluorophores are selected. Keep this setting for a full dye scouting run or uncheck any dye, which you want to rule out for your screen. With default parameter settings, the measurement time is around 20 min per dye.
TIP
A high analyte concentration is recommended to ensure saturated association signals (if possible, 10-fold higher than the expected KD value). A full dissociation is not required for this assay as the aim of this screening is to check for maximal signal amplitudes of the different dyes during association. Select Hit Screening as type of analysis. The software will plot the amplitude signal of the analyte association for each tested dye in a bar diagram. The higher the fluorescent change signal (%), more suitable is the dye to study the interaction. For further questions, please contact the support team at support@dynamic-biosensors.com
Useful Order Numbers
- Table 2. Order Numbers
| Product Name | Comment | Order No |
|---|---|---|
| heli X ® Adapter Chip | Chip with 2 detection spots | ADP-48-2-0 |
| heli X ® Amine Coupling Kit 1 | For five individual conjugation | |
| reactions | HK-NHS-1 |
Contact
Dynamic Biosensors GmbH
- Perchtinger Str. 8/10
- 81379 Munich
- Germany
Dynamic Biosensors, Inc.
- 300 Trade Center, Suite 1400
- Woburn, MA 01801
- USA
- Order Information order@dynamic-biosensors.com
- Technical Support support@dynamic-biosensors.com
- www.dynamic-biosensors.com
- Instruments and chips are engineered and manufactured in Germany.©2024 Dynamic Biosensors GmbH | Dynamic Biosensors, Inc. All rights reserved. [1] TE40: 10 mM Tris, 40 mM NaCl, 0.05 % Tween20, 50 μM EDTA, 50 μM EGTA
- www.dynamic-biosensors.com
References
Read User Manual Online (PDF format)
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