NTA chip creation: Difference between revisions
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Revision as of 18:26, 16 May 2007
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.
