These tools will no longer be maintained as of December 31, 2024. Archived website can be found here. PubMed4Hh GitHub repository can be found here. Contact NLM Customer Service if you have questions.


Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

  • Title: FRET-based Ca2+ measurement in B lymphocyte by flow cytometry and confocal microscopy.
    Author: Adachi T, Tsubata T.
    Journal: Biochem Biophys Res Commun; 2008 Mar 07; 367(2):377-82. PubMed ID: 18167311.
    Abstract:
    Upon the B cell antigen receptor (BCR) ligation Ca(2+) mobilization is induced, which is essential for activation of downstream signaling molecules such as MAP kinase. Although synthetic fluorescent chelators such as Fluo-4 and Indo-1 are widely used for Ca(2+) measurement upon BCR ligation, they are leaked or unfavorably localized into some organelles with time post loading. To solve these problems, we introduce a genetically encoded fluorescent indicator cameleon which is a fluorescence resonance energy transfer (FRET)-based indicator comprising two fluorescent proteins (CFP and YFP) and two Ca(2+)-responsive elements (a variant of calmodulin (CaM) and a CaM-binding peptide). Here, we demonstrate that cameleon as well as a conventional synthetic Ca(2+) indicator enables Ca(2+) measurement by flow cytometry clearly upon BCR ligation. In addition, confocal microscopy analysis allows us to detect cameleon-based Ca(2+) mobilization in a single cell upon BCR ligation.
    [Abstract] [Full Text] [Related] [New Search]