HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Cisar, L. A. Articles by Bensadoun, A. Search for Related Content PubMed PubMed Citation Articles by Cisar, L. A. Articles by Bensadoun, A. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol 26, 380-386, Copyright © 1985 by Lipid Research, Inc.

ARTICLES

Enzyme-linked immunosorbent assay for rat hepatic triglyceride lipase

LA Cisar and A Bensadoun

A noncompetitive enzyme-linked immunosorbent assay to measure rat hepatic triglyceride lipase (H-TGL) was developed. Antibodies to rat H- TGL were purified from goat antisera by immunoadsorption on an H-TGL- Sepharose 4B column. Routinely, Immulon 2 Removawell strips were coated with the purified antibody overnight at 4 degrees C. After blocking the wells with bovine serum albumin (BSA) for 2 hr at room temperature, standards (0.85 ng/ml-13.1 ng/ml) or samples were added to the wells and were incubated with the bound anti-rat H-TGL overnight at 4 degrees C. The standards and samples had been pretreated with 5-20 mM SDS for 30 min at room temperature and were then diluted so that the final SDS concentration in the assay was 1 mM or less. The pretreatment with SDS was necessary to achieve maximal immunoreactivity. The sample incubation was followed by an overnight incubation at 4 degrees C with an anti-rat H-TGL-horseradish peroxidase conjugate. Rat H-TGL was detected by the color development after the addition of 0.4 mg/ml of o- phenylenediamine in 0.01% H2O2, 0.1 M citrate phosphate, pH 5.0. A linear relationship was obtained between absorbance at 490 nm and the amount of highly purified rat H-TGL used as a standard. Inclusion of 1 M NaCl in the assay buffer (1% BSA, 0.05% Tween 20, 10 mM phosphate, pH 7.4) during the sample and conjugate incubations minimized non-specific interactions. Recoveries of purified rat H-TGL added to a rat liver perfusate sample ranged from 98.6% to 103%.(ABSTRACT TRUNCATED AT 250 WORDS)
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				H. Gonzalez-Navarro, Z. Nong, M. J. A. Amar, R. D. Shamburek, J. Najib-Fruchart, B. J. Paigen, H. B. Brewer Jr., and S. Santamarina-Fojo The Ligand-binding Function of Hepatic Lipase Modulates the Development of Atherosclerosis in Transgenic Mice J. Biol. Chem., October 29, 2004; 279(44): 45312 - 45321. [Abstract] [Full Text] [PDF]

				M. Verges, A. Bensadoun, J. Herz, J. D. Belcher, and R. J. Havel Endocytosis of Hepatic Lipase and Lipoprotein Lipase into Rat Liver Hepatocytes in Vivo Is Mediated by the Low Density Lipoprotein Receptor-related Protein J. Biol. Chem., March 5, 2004; 279(10): 9030 - 9036. [Abstract] [Full Text] [PDF]

				H. Gonzalez-Navarro, Z. Nong, L. Freeman, A. Bensadoun, K. Peterson, and S. Santamarina-Fojo Identification of mouse and human macrophages as a site of synthesis of hepatic lipase J. Lipid Res., May 1, 2002; 43(5): 671 - 675. [Abstract] [Full Text] [PDF]

				R. A. Sendak, D. E. Berryman, G. Gellman, K. Melford, and A. Bensadoun Binding of hepatic lipase to heparin: identification of specific heparin-binding residues in two distinct positive charge clusters J. Lipid Res., February 1, 2000; 41(2): 260 - 268. [Abstract] [Full Text]

				B. J. M. van Vlijmen, A. Rohlmann, S. T. Page, A. Bensadoun, I. S. T. Bos, T. J. C. van Berkel, L. M. Havekes, and J. Herz An Extrahepatic Receptor-associated Protein-sensitive Mechanism Is Involved in the Metabolism of Triglyceride-rich Lipoproteins J. Biol. Chem., December 3, 1999; 274(49): 35219 - 35226. [Abstract] [Full Text] [PDF]

				N. Bergeron, L. Kotite, M. Verges, P. Blanche, R. L. Hamilton, R. M. Krauss, A. Bensadoun, and R. J. Havel Lamellar lipoproteins uniquely contribute to hyperlipidemia in mice doubly deficient in apolipoprotein E and hepatic lipase PNAS, December 22, 1998; 95(26): 15647 - 15652. [Abstract] [Full Text] [PDF]