Most used basic analysis running buffer
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 a working buffer for intermolecular binding experiments.
This buffer is filtered.
150 mM NaCl
10 mM HEPES
0.005% T20 surfactants
Amine coupling reagent for COOH functionalized sensor chip
Amine coupling is a method of covalently coupling protein amines to the COOH sensor chip when the ligand is a protein. Since the protein is rich in amino acids with amine residues such as lysine and arginine, it can be very successfully immobilized on the COOH sensor chip.
Since this immobilized ligand is covalently bonded, it is not desorbed during the regeneration step and has the advantage that it can be used continuously as long as the protein activity is not reduced.
However, since the protein active site is fixed on the surface of the sensor chip and the activity is sometimes reduced, if no signal is seen in the positive sample, another immobilization method (if the protein has a tag, the protein can be directionally fixed by non-covalent specific binding). We recommend that you choose
Amine coupling kit
Activation, blocking, and elution buffers for ligand immobilization are included. This reagent set can be used approximately 100 times.
Activation reagent 1: Activation buffer (NHS, 25 ml)
Activation reagent 2: EDC, 1 g
Acetate buffer 25 ml
Blocking reagent: Quenching buffer, 30 ml
Borate elution buffer, 50 ml
Applied sensor chip
ligand immobilization buffer
This buffer is pH-specific acetate buffer for ligand immobilization . When this buffer is used with the amine coupling kit, the ligand can be effectively immobilized on the COOH functionalized chip.
Due to the different pI values of the ligand protein, a wide range of acetate buffer is available to determine the optimal pH at which the protein immobilization used is well controlled . It is recommended to do
5mM acetate buffer pH4.0, 50ml
5mM acetate buffer pH4.5, 50ml
5mM acetate buffer pH 5.0, 50 ml
5mM acetate buffer pH5.5, 50ml
Reagents related to NTA-Histag binding
Recombinant proteins often have his-tags. If the protein is not immobilized well by the amine coupling method or the binding signal is weak, a method of non-covalently fixing the his-tag site to the NTA sensor chip can be selected.
This method is useful when the ligand protein is diverse because the ligand protein is also desorbed during the regeneration step. For the same reason, the single cycle kinetics method is recommended for kinetics evaluation.
Since the bond between NTA and 6xhistidine is not very strong, it may be continuously desorbed during the experiment. After sufficient washing, it is recommended to proceed with the analysis when drift is minimized.
The NiHC1000M sensor chip provided by iClubio is a special product that strongly improves the binding force between NTA and 6xhistidine. If you are concerned about the above issues, we recommend using NiHC1000M.
Capture kit for NTA functionalized sensor chip
About 100 times for immobilization of Histag protein.
Running buffer, 50ml
Activation reagent: Nikel ion solution, 50ml
EDTA regeneration buffer, 50ml
Imidazole regeneration buffer, 50ml
Applied sensor chip
These are the main regeneration reagents that can regenerate the ligand-immobilized sensor chip by desorbing the analyte bound to the ligand.
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
Reagents for device tubing care
Desorb reagent for maintenance/maintenance which can decontaminate device tubing.
Desorb solution1: 0.5% SDS, 500ml
Desorb solution2: 50mM Glycine pH9.5, 500ml