Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4286

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Harlow, E.
Right arrow Articles by Lane, D.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Harlow, E.
Right arrow Articles by Lane, D.
Related Collections
Right arrow Antibodies, general
Right arrow Handling Antibodies, general
Right arrow Labeling Antibodies
Right arrow Immunology, general
Right arrowRelated Protocols
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
Legend icon

protocolProtocol

Labeling Antibodies with Biotin

Ed Harlow and David Lane

This protocol was adapted from "Handling Antibodies," Chapter 4, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

The first 15% of the full text of this article appears below.


INTRODUCTION

This protocol describes the covalent coupling of antibodies to biotin. Biotin groups bind with extremely high affinity to streptavidin and avidin, both of which are available commercially coupled with enzymes, fluorescent dyes, or iodine. A biotinylated primary antibody, therefore, can be detected with any of a wide variety of different labels. The biotinylation reaction is simple and mild, and rarely inactivates the antibody. Biotinylated antibodies can be stored for years without loss of activity.


MATERIALS

Reagents

recipe caution Dimethyl sulfoxide (DMSO, anhydrous)

N-Hydroxisuccinimide . . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?

Related Protocols

Antibody Purification on Protein A or Protein G Columns
Ed Harlow and David Lane
CSH Protocols 2006: 4283. [Extract] [Full Text]

Purification of Antibodies on an Antigen Column
Ed Harlow and David Lane
CSH Protocols 2006: 4284. [Extract] [Full Text]

Labeling Monoclonal Antibodies by Biosynthesis
Ed Harlow and David Lane
CSH Protocols 2006: 4288. [Extract] [Full Text]