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Molecular Interaction Analysis Tools
Amine coupling consumables
Amine coupling is a method of covalently binding the amine of the protein to the COOH sensor chip. Since proteins are rich in amino acids containing amine residues such as lysine and arginine, they can be very successfully immobilized on a COOH sensor chip. Since the ligand fixed in this way is covalently bonded, it does not detach during the regeneration stage and has the advantage of being able to continue to be used as long as protein activity is not reduced. However, since the protein active site may be fixed to the surface of the sensor chip and the activity may be reduced, if no signal is visible even in the positive sample, use another immobilization method.
Amine coupling kit
Contains activation, blocking, and elution buffers for ligand immobilization. This reagent set can be used approximately 100 times.
Contents
Activation reagent 1: Activation buffer (NHS, 25 ml)
Activation reagent 2: EDC, 1g
Acetate buffer 25ml
Blocking reagent: Quenching buffer, 30 ml
Borate elution buffer, 50 ml
Running buffer, 50ml
Applicable sensor chips
COOH-Au chip, C-Dex100, HC1000M
Immobilization buffer
This buffer is a pH-specific acetate buffer for immobilization of ligands. Using this buffer with an amine coupling kit can effectively immobilize the ligand onto the COOH chip. Because the pI values of the ligand proteins are different, it is recommended to have a wide range of acetate buffers to determine the optimal pH.
Product list
5mM acetate buffer pH4.0, 50ml
5mM acetate buffer pH4.5, 50ml
5mM acetate buffer pH5.0, 50ml
5mM acetate buffer pH5.5, 50ml
Histag capture kit
If the protein is not well immobilized by the amine coupling method or the binding signal is weak, you can select the method of immobilizing the His-tag on the NTA sensor chip. This method is good to use when the ligand proteins are diverse because even the ligand proteins are desorbed during the regeneration step. For the same reason, the single-cycle kinetics method is recommended when performing kinetics evaluation. Since the bond between NTA and 6xhistidine is not a very strong bond, it may continue to desorb during the experiment. It is recommended that analysis be performed when drift is minimized after sufficient washing. The NiHC1000 sensor chip provided by icluebio is a special product that greatly improves the binding force between NTA and 6xhistidine.
Histag capture kit
About 100 cycles for immobilization of Histag protein
This is a standard reagent set for use.
Lists
Running buffer, 50ml
Activation reagent: Nikel ion solution, 50ml
EDTA regeneration buffer, 50ml
Regeneration buffer, 50ml
Applicable sensor chips
NTA-Au chip
NiD200M
NiHC1000M
NEW
NTA Regeneration kit
EDTA or NaCl solutions are generally used to regenerate the sensor chip after completing the experiment. However, when the ligand immobilization level is high, over 5000RU, it is often difficult to completely restore the sensor chip to its initial state with a general regeneration solution. In this case, the sensor chip can be completely restored by using the NTA Regeneration kit optimized by icluebio.
This is a set of reagents for approximately 100 uses for the regeneration of NTA sensor chips.
Lists
NTA Regeneration solution 1, 25ml
NTA Regeneration solution 2, 25ml
NTA Regeneration solution 3, 25ml
Applicable sensor chips
NTA-Au chip
NiD200M
NiHC1000M
Biotin-molecule
capture kit 1
(for COOH chips)
Avidin-biotin interaction is based on the strong affinity bond between avidin and biotin. Avidin is a protein that is rich in amino acids containing amine residues, so it can be covalently linked to the COOH sensor chip using the amine coupling method. Biotin-molecules capture kit 1 (BCK1) consists of amine coupling-based reagents to covalently bind Neutravidn, a type of avidin, to COOH sensor chips (PCCH1000, DCCH1100, HC1000). After immobilizing Neutravidin on the sensor chip using BCK1 and the Amine coupling kit, you can easily and successfully immobilize your biotinylated molecules (protein, aptamer, peptides, etc.).
Biotin-molecules capture kit 1 (for COOH chip)
This is a set of reagents for approximately 10 uses for immobilization of biotinylated molecules. Must be used together with an amine coupling kit.
Contents
Neutravidin solution, 10ul x 5ea
Running buffer, 50ml
Immobilization buffer, 1ml x 5ea
Regeneration buffer, 25ml
Regeneration buffer, 50ml
Applicable sensor chips
COOH-Au chip
C-Dex100
HC1000
NEW
Biotin-molecule
capture kit 2
(for Biotin chips)
Avidin can bind up to four biotin molecules, so the ligand can be fixed to the sensor chip using a sandwich method using the biotin of the sensor chip and the biotin of the ligand. Biotin-molecules capture kit 2 (BCK2) consists of reagents to immobilize biotinylated molecules on a biotin sensor chip in a sandwich manner. Since the ligand-bound through the BCK2 reagent is very strongly bound based on the strong affinity bond of avidin-biotin, it is not desorbed by general regeneration solutions such as glycine, and analysis can proceed as if it were a covalent bond. In addition, after the experiment is completed, the biotin sensor chip can be regenerated to its initial state through the optimized regeneration solutions included in BCK2.
Biotin-molecules capture kit 2 (for Biotin chip)
This is a set of reagents for approximately 10 uses for immobilization of biotinylated molecules.
Contents
Neutravidin solution, 10ul x 5ea
Running buffer, 50ml
Regeneration buffer, 25ml
Reuse solution 1, 25ml
Reuse solution 2, 25ml
Reuse solution 3, 25ml
Applicable sensor chips
Biotin-Au chip
Buffers and Reagents
Running buffers
10X HBST buffer (50ml, 500ml)
Hepes buffered saline with tween20
It is a 10-fold concentrated HBST buffer. Ligand immobilization (amime coupling, NTA-His, Avidin-biotin), It is used as an operating buffer for intermolecular binding experiments. This buffer is filtered. Depending on the experiment, you can choose a running buffer with various compositions. In addition, you can select a PBS-type running buffer.
ingredients
150mM NaCl
10 mM HEPES
0.005% T20 surfactants
pH7.4
Regeneration buffers
These are major regeneration reagents that can be regenerated as a ligand-immobilized sensor chip by detaching the analyte bound to the ligand.
product list
10mM Glycine-HCl pH1.5, 50ml
10mM Glycine-HCl pH2.0, 50ml
10mM Glycine-HCl pH2.5, 50ml
10mM Glycine-HCl pH3.0, 50ml
50 mM NaOH, 50 ml
0.5% SDS, 50ml
Desorb solutions
Desorb reagent for maintenance/management that can remove contaminants from device tubing.
product list
Desorb solution1: 0.5% SDS, 500ml
Desorb solution2: 50mM Glycine pH9.5, 500ml
