|
|
||||||||
Correspondence to:
Mark H. Doolittle.
This manuscript describes the problems inherent in combining immunoprecipitation of lipoprotein lipase (LPL) with its detection by Western blot, and how these problems can be circumvented by the preparation of suitable immunoreagents. These reagents used during the immunoprecipitation step, include Fab fragments of the primary antibody (chicken anti-bovine LPL), and a covalently linked immunomatrix of the secondary antibody (rabbit anti-chicken IgG). The use of these reagents in conjunction with Western blot detection virtually eliminates the problem of non-relevant protein detection when analyzing LPL from complex biological samples. Moreover, this approach can be adapted to detect any protein with the same inherent problems as LPL, such as hepatic lipase.Doolittle, M. H., O. Ben-Zeev, and V. Briquet-Laugier. Enhanced detection of lipoprotein lipase by combining immunoprecipitation with Western blot analysis. J. Lipid Res. 1998. 39: 934942.
Supplementary key words:
affinity purified chicken anti-LPL antibody, chicken IgG Fab production, immunomatrix
Copyright © 1998 by Lipid Research, Inc.
Paper on Methodology
Enhanced detection of lipoprotein lipase by combining immunoprecipitation with Western blot analysis
Mark H. Doolittlea,
Osnat Ben-Zeeva, and
Véronique Briquet-Laugiera
a Lipid Research Laboratory, West Los Angeles VA Medical Center, and Department of Medicine, University of California-Los Angeles, Los Angeles, CA 90073
![]()
CiteULike
Complore
Connotea
Del.icio.us
Digg
Reddit
Technorati What's this?
This article has been cited by other articles:
![]() |
O. Ben-Zeev and M. H. Doolittle Maturation of Hepatic Lipase: FORMATION OF FUNCTIONAL ENZYME IN THE ENDOPLASMIC RETICULUM IS THE RATE-LIMITING STEP IN ITS SECRETION J. Biol. Chem., February 13, 2004; 279(7): 6171 - 6181. [Abstract] [Full Text] [PDF] |
||||
![]() |
L. Zhang, G. Wu, C. G. Tate, A. Lookene, and G. Olivecrona Calreticulin Promotes Folding/Dimerization of Human Lipoprotein Lipase Expressed in Insect Cells (Sf21) J. Biol. Chem., August 1, 2003; 278(31): 29344 - 29351. [Abstract] [Full Text] [PDF] |
||||
![]() |
O. Ben-Zeev, H. Z. Mao, and M. H. Doolittle Maturation of Lipoprotein Lipase in the Endoplasmic Reticulum. CONCURRENT FORMATION OF FUNCTIONAL DIMERS AND INACTIVE AGGREGATES J. Biol. Chem., March 15, 2002; 277(12): 10727 - 10738. [Abstract] [Full Text] [PDF] |
||||
![]() |
V. Briquet-Laugier, O. Ben-Zeev, A. White, and M. H. Doolittle cld and lec23 are disparate mutations that affect maturation of lipoprotein lipase in the endoplasmic reticulum J. Lipid Res., November 1, 1999; 40(11): 2044 - 2058. [Abstract] [Full Text] |
||||
![]() |
W.-J. Shen, K. Sridhar, D. A. Bernlohr, and F. B. Kraemer Interaction of rat hormone-sensitive lipase with adipocyte lipid-binding protein PNAS, May 11, 1999; 96(10): 5528 - 5532. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Journal of Biological Chemistry |
| Molecular and Cellular Proteomics | ASBMB Today |