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Labeling Protein with Thiol-reactive Probes

QY Qingrong Yan

Published: Jun 5, 2011 DOI: 10.21769/BioProtoc.82 Views: 12549

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Abstract

This protocol is used to label protein or peptide with a maleinmide or iodoacetamide conjugated fluorescent probe through the free cysteine.

Materials and Reagents

  1. Thiol-reactive Maleimide probes (Life Technologies, Invitrogen™)
    Note: A large collection of probes are available from Invitrogen.
  2. Tris-(2-carboxyethyl)phosphine (TCEP) (Life Technologies, Invitrogen™, catalog number: T2556 )
  3. Dithiothreitol (DTT) (Life Technologies, Molecular Probes®, catalog number: D1532 )
  4. DMSO
  5. Dissolving buffer (see Recipes)

Equipment

  1. Aluminum foil
  2. Sephadex G-25 column

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Biochemistry > Protein > Labeling

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1 Q&A

One question: In some protocols (very similar to yours), it's...
edit 1 Answer 1 View Jun 27, 2011