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

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

Journal of Lipid Research, Vol. 43, 167-173, January 2002
Copyright © 2002 by Lipid Research, Inc.

Methods

Sandwich immunoassay for the measurement of murine syndecan-4

Correspondence to: André Bensadoun, To whom correspondence should be addressed., ab55{at}cornell.edu (E-mail)

A sensitive and specific immunoassay for mouse syndecan-4 is described. The assay is a non-competitive direct sandwich enzyme-linked immunosorbent assay based on the production of an affinity-purified polyclonal antibody directed against syndecan-4 extracellular domain, used both as a trapping and detecting antibody. The standard curve is constructed with recombinant 6-His-tagged syndecan-4 extracellular domain. The assay allowed quantification of syndecan-4 core protein in the partially purified proteoglycan fraction from adipocyte 3T3-F442A cells, as well as in the cellular whole and clarified lysates. Removal of glycosaminoglycan chains on syndecan-4 core protein is required for maximal epitope exposure. The standard curve ranged between 7 and 70 fmol per well. For the 14 standard curves run on different days, the absorbance at 490 nm for 35.5 fmol of recombinant syndecan-4 was 0.610 ± 0.110 (n = 14) with a corresponding blank absorbance of 0.089 ± 0.030. At low (5.7 fmol) and high (42.8 fmol) levels of whole cell syndecan-4, the intra-assay and inter-assay coefficients of variation were 4.9 and 15.3 percent, and 3.3 and 9.7 percent, respectively.

A survey of the syndecan-4 expression in various mouse tissues shows that syndecan-4 is highly expressed in the kidney, brain, testes and liver, but can also be measured in several different adipose tissue sites. — Rioux, R., R. Y. Landry, and A. Bensadoun. Sandwich immunoassay for the measurement of murine syndecan-4. J. Lipid Res. 2002. 43: 167–173.

Supplementary key words: enzyme-linked immunosorbent assay, quantification, syndecan-4, heparan sulfate proteoglycans, adipose tissue

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. A. Julien, P. Wang, C. A. Haller, J. Wen, and E. L. Chaikof Mechanical strain regulates syndecan-4 expression and shedding in smooth muscle cells through differential activation of MAP kinase signaling pathways Am J Physiol Cell Physiol, January 1, 2007; 292(1): C517 - C525. [Abstract] [Full Text] [PDF]

				A. Schmidt, F. Echtermeyer, A. Alozie, K. Brands, and E. Buddecke Plasmin- and Thrombin-accelerated Shedding of Syndecan-4 Ectodomain Generates Cleavage Sites at Lys114-Arg115 and Lys129-Val130 Bonds J. Biol. Chem., October 14, 2005; 280(41): 34441 - 34446. [Abstract] [Full Text] [PDF]