| Chapter title |
High-throughput biotinylation of proteins.
|
|---|---|
| Chapter number | 13 |
| Book title |
High Throughput Protein Expression and Purification
|
| Published in |
Methods in molecular biology, January 2009
|
| DOI | 10.1007/978-1-59745-196-3_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-58829-879-9, 978-1-59745-196-3
|
| Authors |
Kay, Brian K, Thai, Sang, Volgina, Veronica V, Brian K. Kay, Sang Thai, Veronica V. Volgina, Kay, Brian K., Volgina, Veronica V. |
| Abstract |
One of the more useful tags for a protein in biochemical experiments is biotin, because of its femtomolar dissociation constant with streptavidin or avidin. Robust methodologies have been developed for other the in vivo addition of a single biotin to recombinant protein or the in vitro enzymatic or chemical addition of biotin to a protein. Such modified proteins can be used in a variety of experiments, such as affinity selection of phage-displayed peptides or antibodies, pull-down of interacting proteins from cell lysates, or displaying proteins on arrays. We present three complementary approaches for biotinylating proteins in vivo in Escherichia coli or in vitro using chemical or enzymatical reactions all of which can be scaled up to tag large numbers of proteins in parallel. |
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