Forum :: Adding Neutravidin to CM5 chip

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These are the posts of the old forum. It was not possible to transfer the user data, so they are missing in most of the posts. For new questions, go to the general discussions.

TOPIC: Adding Neutravidin to CM5 chip

Adding Neutravidin to CM5 chip 11 Feb 2013 01:00 #1

I would like to couple a CM5 chip with Neutravidin, block with Ethanolamine and then add my biotinylated protein (looking at protein-peptide binding). Has anyone had experience with this setup? I guess I am interested in knowing how well it worked for others and what amount of Neutravidin they have had good luck with.

Any other hints/suggestions would be greatly appreciated.

Claude Bessy

Adding Neutravidin to CM5 chip 11 Feb 2013 01:00 #2

Hi Claude

I don't have a perfect answer for you but some references to start with.

I would start with a normal amine immobilization and culculate what desnity you want.
As an alternative you can use the SA-sensorchip with straptavidin on the surface. That will speed up your experiment.
You can also use the CAP-regaent sensor chip. This is a more flexible way of immobilization using the biotin-avidin system. Take a look!

O'Shannessy, D. J., Brigham-Burke, M., and Peck, K.; Immobilization chemistries suitable for use in the BIAcore surface plasmon resonance detector. Anal.Biochem. (205): 132-136; 1992.
Gitlin, G., Bayer, E. A., and Wilchek, M.; Studies on the biotin-binding sites of avidin and streptavidin. Tyrosine residues are involved in the binding site. Biochem.J. (269): 527-530; 1990.

Wilchek, M. and Bayer, E. A.; Introduction to avidin-biotin technology. Methods Enzymol. (184:5-13.): 5-13; 1990.

best regards

The following user(s) said Thank You: Maedehmozneb

Adding Neutravidin to CM5 chip 11 Feb 2013 01:00 #3

Thanks Arnoud!

Sounds like this is going to be very experimental. I was hoping to reduce costs by manually adding the avidin (I know, this will probably haunt me in the end).

I will post my findings for someone to use in the future.