dynamic BIOSENSORS NOR-Ga heliXcyto RT-IC Measurements in the Green Channel Instruction Manual

: June 1, 2024
: dynamic BIOSENSORS

Table of Contents

dynamic BIOSENSORS NOR-Ga heliXcyto RT-IC Measurements in the Green
Specifications
Product Description
FAQ
Key Features
Product Description
Preparation
Application Note
Contact
References
Read User Manual Online (PDF format)
Download This Manual (PDF format)

dynamic BIOSENSORS NOR-Ga heliXcyto RT-IC Measurements in the Green

Channel

Specifications

Product Name: heliXcyto Normalization Solution (Green Dye)
Model: NOR-Ga v1.0
Order Number: NOR-Ga
For Research Use Only

Product Description

The heliXcyto Normalization Solution (Green Dye) is designed for RT-IC measurements in the green channel. It is used to ensure accurate and reliable fluorescent signal normalization during experiments.

Key Features

Green dye for normalization
Limited shelf life (refer to expiry date on label)

Preparation

The concentration of the normalization solution should match the highest signal from the analyte. Excitation power should be adjusted based on fluorophore concentration, expected binding signal, and chip type.

Application Note

In RT-IC measurements, ensuring the normalization solution’s fluorescent signal matches the analyte’s signal is crucial for accurate results. The table below provides guidance on normalization solution concentration, analyte dye concentration, and excitation power suitable for heliXcyto M5 chip.

Analyte Dye Conc.	Excitation Power	Normalization Solution Conc.	Dilution
25 nM	0.5	25 nM	1:400

Note: Optimization may be required based on system specifics for final signal recording.

Contact Information

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

FAQ

Q: What is the shelf life of the heliXcyto Normalization Solution?

A: The product has a limited shelf life, please refer to the expiry date on the label for specific information.

Normalization solution (green dye)

for RT-IC measurements in the green channel Dynamic Biosensors GmbH & Inc. NOR-Ga v1.0

Key Features

For the normalization of the fluorescent signals on Spot 1 and Spot 2 of a heliXcyto chip
Enables correct real-time referencing of the green fluorescent signals during RT-IC measurements
Compatible with all chips
The Normalization solution (green dye) contains a hydrophilic green dye with a single negative net charge

Product Description

Order Number: NOR-Ga
Table 1. Contents and Storage Information

For research use only.

This product has a limited shelf life, please see expiry date on label.

Preparation

Use this green dye normalization solution for RT-IC measurements in the green channel (analyte label-dependent). Dilute the 10 μM normalization stock solution to a working concentration with running buffer.
The concentration of the normalization solution should approximately correspond to the fluorophore concentration in the highest analyte concentration to be measured. This can be calculated using following equation:

Concentration of the normalization solution in the desired color
Concentration of dye in the labeled analyte solution
Highest concentration of analyte that should be measured
DOL : Degree of labelling (ratio of dye to analyte)

Diluted solutions can be stored at 2-8°C for up to 7 days.

Application Note

In the RT-IC measurement, the fluorescent signal of the normalization solution should be in a similar range as the highest signal coming from bound analyte (raw data). The absolute fluorescent signal is dependent on normalization solution concentration and the excitation power applied in the measurement. The excitation power has to be selected based on the following parameter:

a. Fluorophore concentration in analyte solution:
The fluorophore concentration depends on the analyte concentration used in the measurement as well as the degree of labeling of the analyte. For high DOL and high analyte concentrations, lowering the excitation power might be required.
b. Expected binding signal:
Highly expressed targets on a cell can bind more molecules of labeled analyte. In case of highly overexpressed targets a strong binding signal can be expected. To avoid the shutter closing, lowering the excitation power might be considered.
c. Chip type:
Different chip types have varying fluorescent backgrounds. The bigger the traps and the more traps on the chip, the higher the background signal. Therefore, L5 chips might require lower excitation power than applied to M5 chips.

For a starting point of excitation power and norm. solution concentration to be used in an RT-IC experiment, please refer to Table 2.

Table 2. Relation of fluorophore concentration, normalization solution concentration, and excitation power suitable for a M5 chip

Note: This table is for your guidance. However, the final signal recorded in the depends on many factors. Thus, some optimization will be required for each system.