Isothiocyanobenzyl-NTAI279 Product Manual

Isothiocyanobenzyl-NTA* (ITC-Bz-NTA) is used for the introduction of a chelating function to various materials such as amine compound, biomolecules, and surfaces through their amino groups under physiological conditions. Since NTA makes a stable chelate with heavy metal ions, NTA-conjugated compound, biomolecules, and surfaces can hold heavy metals. Such heavy metal ion chelate can be used for detection or separation of specific substances that interact with heavy metal ions.
*NTA : Nitrilotriacetic acid

Store at -20℃.

ImmunoglobulinG (IgG)

1. Prepare a 1 mg/ml IgG solution with 100 mmol/l Bicine Buffer (pH 8.5).
2. Dissolve 1 mg ITC-Bz-NTA with 50 μl DMSO.
   *The DMSO solution is stable for 1 week at -20℃.
3. 100 μl IgG solution and 10 μl ITC-Bz-NTA solution in a microtube.
4. Incubate the mixture at 37℃ for 1 hour.
5. Purify the NTA-conjugate with a gel filtration or a dialysis.

 

DNA oligomer with an amino group

1. Prepare a DNA oligomer solution with 100 mmol/l Bicine buffer(pH 8.5).
2. Dissolve 1 mg ITC-Bz-NTA with 50 μmol/l DMSO*.
   • The DMSO solution is stable for 1 week at -20°C.
3. Mix the DNA oligomer solution and ITC-Bz-NTA solution in a microtube ^a).
4. Incubate the mixture at 37℃ for 1 hour. 5. Purity the NTA-conjugated with a gel filtration.

a) Generally, 20 times equivalent of ITC-Bz-NTA should be added to the DNA oligomer.
The volume of the DMSO solution should be less than 20% of the DNA oligomer solution.

Preparation of Ni-NTA complex

1. Prepare NiCl₂ solution with 10 mmol/l HCl.
2. Add the 5 equivalent of Ni solution.
3. Adjust the pH at 7 with the 0.1 mol/l NaHCO₃.

An appropriate number of NTA to be introduced will differ optimize the protocol to prepare a suitable NTA-conjugated materials.