NTA chip creation: Difference between revisions
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Talk to[[User_talk:Jfuller| Jason Fuller]] to discuss this protocol. | Talk to[[User_talk:Jfuller| Jason Fuller]] to discuss this protocol. | ||
==References== | |||
<biblio> | |||
#SPRNTAuse pmid=9344407 | |||
</biblio> | |||
<!-- You can tag this protocol with various categories. See the [[Categories]] page for more information. --> | <!-- You can tag this protocol with various categories. See the [[Categories]] page for more information. --> | ||
[[Category:Protocol]] | [[Category:Protocol]] | ||
Latest revision as of 15:42, 21 March 2008
Overview
The purpose of this protocol is to provide a detailed method for the creation of an NTA surface on top of a carboxymethylated dextran chip. This protocol specifically focuses on the BiaCore CM5 chip, but could be adapted to other similar chips.
Materials
- CM5 Chip (New)
- EDC (50µl)
- NHS (50µl)
- Wash Bottle with Deionized Water
- 3mg/ml NTA-NH2 (100µl)
Definitions
- EDC: 1-Ethyl-3-(dimethylaminopropyl)cabodiimide HCL
- NHS: N-Hydroxysuccinimide
- NTA-NH2: (S)-N-(5-Amino-1-Carboxypentyl)iminodiacetic acid, pH adjusted to pH 8.0
Procedure
- To the gold side (concave) of a CM5 chip mix 50μl each EDC and NHS.
- Incubate for 20 minutes
- Wash chip with DI and then dry with house air.
- Add 100 ul of 3mg/ml NTA-NH2 to the chip
- Incubate for 30 minutes.
- Wash chip with DI and then dry with house air.
- Install into the BiaCore and perform NTA Surface Preparation.
Contact
Talk to Jason Fuller to discuss this protocol.
References
- Nieba L, Nieba-Axmann SE, Persson A, Hämäläinen M, Edebratt F, Hansson A, Lidholm J, Magnusson K, Karlsson AF, and Plückthun A. BIACORE analysis of histidine-tagged proteins using a chelating NTA sensor chip. Anal Biochem. 1997 Oct 15;252(2):217-28. DOI:10.1006/abio.1997.2326 |
