Purification of the T cell antigen receptor and analysis by blue-native PAGE.

Mahima Swamy, Wolfgang W.A. Schamel

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

The T cell antigen receptor (TCR) is a multi-protein complex composed of six different transmembrane subunits, which form complexes of various sizes on the surface of resting T cells. The stoichiometry of the smallest form was recently determined to be αβγεδεζζ, whereas that of the larger forms is unknown. The roles of the different forms and their ratios are poorly defined. Biochemical analyses to address these questions must focus on the detergent and the best native conditions to maintain the integrity of the complexes. Blue-native polyacrylamide gel electrophoresis (BN-PAGE) is a high-resolution native protein separation method that relies on the dye Coomassie blue to confer negative charge for separation. Using this powerful approach, the size, subunit composition and the relative abundance of the different TCR forms can be studied. We present here four methods to isolate the TCR in a native form and details to analyse it by BN-PAGE.

Original languageEnglish
Title of host publicationT Cell Protocols
EditorsGennaro De Libero
PublisherSpringer
Chapter10
Pages135-150
Number of pages16
Volume514
Edition2nd
ISBN (Electronic) 978-1-60327-527-9
ISBN (Print)978-1-58829-587-3
DOIs
Publication statusPublished - 2009

Publication series

NameMethods in Molecular Biology (Clifton, N.J.)
PublisherSpringer Verlag
ISSN (Print)1064-3745

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    Swamy, M., & Schamel, W. W. A. (2009). Purification of the T cell antigen receptor and analysis by blue-native PAGE. In G. De Libero (Ed.), T Cell Protocols (2nd ed., Vol. 514, pp. 135-150). (Methods in Molecular Biology (Clifton, N.J.)). Springer . https://doi.org/10.1007/978-1-60327-527-9_10