Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



Table of Contents

Expand All | Contract All

Labeling Antibodies Using NHS-Fluorescein

(Protocol summary only for purposes of this preview site)

N-Hydroxysuccinimide (NHS)ester derivatives are among the most commonly used reagents for labeling proteins. The method below can be adapted to use practically any NHS fluorophore. A typical molar excess used in the reactions is 515 times. The actual amounts used will depend on the molecular weight of the dye. Be sure that the antibody to be labeled is at 110 mg/mL and in a buffer at a pH between 7 and 9. Amine-containing buffers (e.g., TRIS) are not compatible with this procedure and must be exchanged (PBS, carbonate, HEPES, and borate are all acceptable).

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

Search for information about other protocols included in the book: