Hi Dear researchers
We want to immobilize enzyme on amino-functionalized silica magnetic nanoparticles (MNP-NH2) using glutaraldehyde (GA) as a cross-linker agent. We used 50 ml of 1% GA solution (in phosphate buffer, pH=7) and 0.5 g amino-functionalized silica magnetic nanoparticles to obtain MNP-NH2-GA. After that 100 mg of MNP-NH2-GA was mixed with 50 mg enzyme in (in phosphate buffer, pH=7) and stirred for 16 h at room temperature. We do all stages based on the published articles but was not observed any changes in Bradford's assay between supernatant and the initial enzyme solutions. Do you have any idea about my problem? What do you think? What is the problem?
I will be grateful for your response,
I used glutaraldehyde for attachment aptamer on the glassy carbon electrode. I hung my electrode vertically above the container with a small hole while the container filled with glutaraldehyde. The container put in the heater with a degree around 40 C. Glutaraldehyde evaporated and covered the electrode surface. Then dropcasted my aptamer on the surface of the electrode. One times that I washed my electrodes before aptamer attachment. The aptamer did not attach on the surface. You can apply other chemistry like EDC/NHS coupling.
You should first make sure that this immobilization method is suitable for your enzyme. Which enzyme are you trying to immobiliz? To the best of my knowledge, many kinds of enzymes might loose their activity during this immobilization approach.
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