NTA chip creation

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Contents

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.

References

  1. Nieba L, Nieba-Axmann SE, Persson A, Hämäläinen M, Edebratt F, Hansson A, Lidholm J, Magnusson K, Karlsson AF, and Plückthun A. . pmid:9344407. PubMed HubMed [SPRNTAuse]
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