Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here to sign up for SAGE Journal Email Alerts today!

Sign In to gain access to subscriptions and/or personal tools.
Journal of Endotoxin Research
This Article
Right arrow Full Text (PDF)
Right arrow References
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Chen, W.H.
Right arrow Articles by Tsai, C-M.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Chen, W.H.
Right arrow Articles by Tsai, C-M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Detection of lipopolysaccharides blotted on nylon membranes

W.H. Chen

Division of Bacterial Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA

P. Balakonis

Division of Bacterial Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA

C-M. Tsai

Division of Bacterial Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA

A sensitive method for the detection of Gram-negative bacterial lipopolysaccharide (LPS) blotted on nylon membranes is described. LPSs are separated by SDS-PAGE and then electrophoretically transferred to nylon membranes. Immobilized LPS is oxidized with periodate and then reacted with a hydrazide conjugated to the steroid, digoxigenin. LPS is visualized by alkaline phosphatase labelled antibodies against the steroid and the enzyme substrate 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium. LPS banding patterns of both rough (R-) and smooth (S-) type LPSs from over 15 different bacterial species are similar to those of silver stained companion gels, but without nonspecific staining of proteins. The detection of S-LPS from Pseudomonas aeruginosa F-D type 1 and R-LPS from Escherichia coli K12 is sensitive to 10-20 ng per lane. The use of this detection system in combination with antibody or lectin studies on identical blots can provide an effective tool in locating the precise position of certain epitopes or sequences in both R- and S-type LPSs on the blots.

Journal of Endotoxin Research, Vol. 2, No. 6, 405-410 (1995)
DOI: 10.1177/096805199600200603
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?