NTA chip creation

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

 * 1) To the gold side (concave) of a CM5 chip mix 50μl each EDC and NHS.
 * 2) Incubate for 20 minutes
 * 3) Wash chip with DI and then dry with house air.
 * 4) Add 100 ul of 3mg/ml NTA-NH2 to the chip
 * 5) Incubate for 30 minutes.
 * 6) Wash chip with DI and then dry with house air.
 * 7) Install into the BiaCore and perform NTA Surface Preparation.

Contact
Talk to Jason Fuller to discuss this protocol.