Journal of Histochemistry and Cytochemistry Priciples for Free Access to Science
  Search:   
    >> Advanced Search

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sippel, T. O.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sippel, T. O.
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  
What's this?

Microfluorometric analysis of protein thiol groups with a coumarinylphenylmaleimide

TO Sippel

Characteristics of thiol adducts formed by the fluorogenic maleimide, N- (4-(7-diethylamino-4-methylcoumarin-3-yl) phenyl) maleimide (CPM), (Sippel: J Histochem Cytochem 29:314, 1981) were determined in solution, on the epithelia in beef cornea paraffin sections, and with an egg white model. Absorption was found maximal at 385-390 nm with a molar absorbancy of not less than 30,000 cm2/mmol, while emission peaked at 465 nm; the spectra and output were hardly affected by hydrolytic opening of the succinimide ring. Fluorescence measured in an epi-illuminating microfluorometer faded rapidly at high magnifications, but the initial output from sections of increasing thickness under a 10 x objective was proportional to the thiol density (concentration x thickness) up to a limit equivalent to an absorbance of nearly 0.5 at 387 nm. The staining included less than 5% nonspecific fluorescence, as determined on duplicate sections blocked by 2,2'-dithiopyridine. Factors affecting the use of CPM for quantitation of both thiols and disulfides are discussed.

Volume 29, Issue 12, pp. 1377-1381, 12/01/1981
Copyright © 1981 by The Histochemical Society


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    What's this?


This article has been cited by other articles:


Home page
 Protein Eng Des SelHome page
M. Taki, M. Shiota, and K. Taira
Transglutaminase-mediated N- and C-terminal fluorescein labeling of a protein can support the native activity of the modified protein
Protein Eng. Des. Sel., February 1, 2004; 17(2): 119 - 126.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
X. Cheng and J. C. Lee
Interactive and Dominant Effects of Residues 128 and 141 on Cyclic Nucleotide and DNA Bindings in Escherichia coli cAMP Receptor Protein
J. Biol. Chem., January 9, 1998; 273(2): 705 - 712.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. M. C. Newton, J. S. Allen, Z. Cao, Z. Qi, X. Jiang, C. Sprencel, J. D. Igo, S. B. Foster, M. A. Payne, and P. E. Klebba
Double mutagenesis of a positive charge cluster in the ligand-binding site of the ferric enterobactin receptor, FepA
PNAS, April 29, 1997; 94(9): 4560 - 4565.
[Abstract] [Full Text] [PDF]



Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 1981

 
Purchase HCS Short Course Manual on HCS site