helix+ User Manual

HIS CAPTURE KIT with red dye Ra

Dynamic Biosensors GmbH & Inc. HK-NTA-1 v4.1

Key Features

This kit is designed for capture of histidine-tagged proteins (His6 or His10) using Tris-NTA.
Compatible with heliX® Adapter Chip.
Includes Adapter strands and Ligand strand modified with Tris-NTA for 20 regenerations.
For functionalization of Spot 1 and Spot 2.
Adapter strands 1 and 2 carry a moderately hydrophilic red dye (Ra) with a single positive net charge.

heliX® Adapter Chip Overview

The heliX® Adapter Chip features two spots for DNA-encoded addressing: Spot 1 for Measurement and Spot 2 for Reference. Each spot has a fixed anchor strand. Spot 1 is functionalized with NTA-Ni and includes exchangeable adapter and ligand strands, along with a dye. Spot 2 serves as a real-time reference and includes exchangeable adapter and ligand-free strands, also with a dye.

Product Description

Order Number: HK-NTA-1

Table 1. Contents and Storage Information

Material	Cap	Concentration	Amount	Buffer	Storage
NTA - Ligand strand	Red	500 nM	2 x 100 µL	TE40 [1]	-20°C
Adapter strand 1 - Ra	Black	400 nM	2 x 100 µL	TE40 [1]	-20°C
Adapter strand 2 - Ra - Ifs	White	200/250 nM	2 x 200 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.

Workflow of a heliX® His - capture assay

The anchor strand (ssDNA) immobilized to the surface of the heliX® Adapter Chip is hybridized with complementary DNA strands modified with Tris-NTA (Nitrilotriacetate).
NTA-DNA surface is activated by loading with divalent nickel ions. His-tagged protein (ligand) is captured on the surface.
Measurement of the analyte binding kinetics or conformational change upon analyte binding.
Surface regeneration either by injection of a high pH solution followed by a new hybridization of fresh DNA-NTA or by simple removal of the his-tagged protein via imidazole wash.

Preparation

Mix 100 µL NTA - Ligand strand with 100 µL Adapter strand 1 - Ra.
Incubate the solution of step 1 at RT at 600 rpm for 30 min to ensure complete hybridization.
Mix 200 µL Adapter strand 2 - Ra - Ifs to the sample after step 2.

The solution (400 µL in total) is ready to use for a biochip functionalization.

Please aliquot and store the ready to use solution at -20°C. Use up within 6 months.

The kit contains material for the preparation of two separate ready to use solutions with 400 µL each.

Assay Setup in heliOS

Go to heliOS > create a New Assay Workflow > add Custom Assay > load His Capture with Kinetics > 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.

TIP: As the stability of his capture is affected by the protein, in case of long dissociations, consider using the conjugation approach.

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
heliX® Adapter Chip	Chip with 2 detection spots	ADP-48-2-0
10x Passivation solution	For passivation of chip surface	SOL-PAS-1-5
Regeneration solution	For regeneration of chip surface	SOL-REG-1-5

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

Website: www.dynamic-biosensors.com

[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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