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P. MATHER, AND GORDON H. SATO VOLUME 199. Cumulative Subject Index Volumes 168–174, 176–194 VOLUME 200. Protein Phosphorylation (Part A: Protein Kinases: Assays, Purification, Antibodies, Functional Analysis, Cloning, and Expression) Edited by TONY HUNTER AND BARTHOLOMEW M. SEFTON VOLUME 201. Protein Phosphorylation (Part B: Analysis of Protein Phosphorylation, Protein Kinase Inhibitors, and Protein Phosphatases) Edited by TONY HUNTER AND BARTHOLOMEW M. SEFTON VOLUME 202. Molecular Design and Modeling: Concepts and Applications (Part A: Proteins, Peptides, and Enzymes) Edited by JOHN J.

We find that 293T cells give a severalfold higher yield of fusion protein than COS cells.

Genet. 32, 601 (1998). 4 Y. Shiio, M. Itoh, and J. Inoue, Methods Enzymol. 254, 497 (1995). 5 P. A. Kolodziej and R. A. Young, Methods Enzymol. 194, 508 (1991). TABLE I COMMONLY USED EPITOPE TAGS Tag Recognized sequence Advantages Disadvantages Original 12CA5 antibody not optimized for use in epitope tagging 9E10 monoclonal may not immunoprecipitate reliably. Endogenous c-myc expression interferes with use as epitope tag Detection by some antibodies requires placement at the protein termini Some commercially available antibodies require additional amino acids to specify recognition HA YPYDVPDYA Highly specific second-generation antibodies available Myc EQKLISEEDL FLAG DYKDDDK Polyhistidine HHHHHH Hybridoma line expressing the 9E10 monoclonal obtainable from the ATCC for use in large-scale projects Epitope easily cleaved off of tagged protein after purification Tagged proteins can be purified on Ni2ϩ affinity matrix.

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