|
Sci. STKE, 27 February 2001 PROTOCOLSImmunoisolation of TCR Signaling Complexes from Jurkat T Leukemic CellsThomas Harder* and Marina Kuhn The Basel Institute for Immunology, Grenzacherstr. 487, CH-4005, Basel, Switzerland. Abstract: The formation of multimolecular assemblies of signaling molecules at the plasma membrane is key to triggering signaling cascades following activation of transmembrane receptors at the plasma membrane. We have developed a method to immunoisolate activated T cell receptors (TCRs) and associated signaling molecules in plasma membrane subdomains from Jurkat T leukemic cells. The immunoisolation procedure for the signaling complexes uses magnetic beads, which are coupled to TCR-activating antibodies. Following mechanical cellular disruption using nitrogen cavitation, conjugates of these beads with Jurkat cells are formed and isolated. TCR-signaling complexes within plasma membrane fragments associate with the antibody-coupled beads and are retrieved along with the beads using a magnet. We found that the immunoisolated plasma membrane fragments are highly enriched for activated TCRs and associated signaling proteins. This isolation procedure allows a detailed and precise biochemical analysis of the assembly of signaling proteins in plasma membrane subdomains. *Corresponding author, E-mail: harder{at}bii.ch.
Citation: T. Harder, M. Kuhn, Immunoisolation of TCR Signaling Complexes from Jurkat T Leukemic Cells. Sci. STKE 2001, pl1 (2001). THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
|
Science Signaling. ISSN 1937-9145 (pre-2008: Science's STKE. ISSN 1525-8882)
Magazine | News | Signaling | Careers | Multimedia | Collections | Help | Site Map | RSS
Subscribe | Feedback | Privacy / Legal | About Us | Advertise With Us | Contact Us
© 2001 American Association for the Advancement of Science. All Rights Reserved.
AAAS is a partner of HINARI, AGORA, PatientInform, CrossRef, and COUNTER.
You have reached the bottom of the page. Back to top