Rapid, antibody-free detection of recombinant proteins on blots using enzyme fragment complementation.
Alternative, antibody-free techniques to western analysis of protein blots can offer reduced assay times for routine analysis of expression of recombinant proteins. We have adapted the commercially available enzyme fragment complementation technology to provide a rapid protein detection method for protein blots based on significantly reducing the number of incubation and washing steps used in traditional approaches, and eliminating the requirement for antibodies. In this article, we highlight the use of this assay for measuring recombinant protein expressed in mammalian cells for a range of applications, including dot blot screening of large numbers of different cell samples, assessment of protein integrity through detection of degradation bands, and characterization of post-translational protein modifications such as glycosylation.
Charter
Neil W
NW
DiscoveRx Corporation, 42501 Albrae St., Fremont, CA, 94538, USA. ncharter@discoverx.com
Horecka
Joe
J
Loh
Chin-Yee
CY
Doan
Albert
A
Wehrman
Tom
T
Olson
Keith R
KR
D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D015153:Blotting, Western / Q000379:methods*; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000032:analysis*
D000906:Antibodies; D011994:Recombinant Proteins