Analysis of B-cell signaling using DT40 B-cell line.

Tomoharu Yasuda, Tadashi Yamamoto

研究成果: Contribution to journalArticle査読

5 被引用数 (Scopus)

抄録

A chicken DT40 B-cell line provides an excellent model system for the analysis of the function of genes related to B-cell antigen receptor (BCR) signaling. Crosslinking of BCR by micro-chain-specific antibody stimulates DT40 cells to undergo apoptosis. Activation of protein- tyrosine kinases (PTKs) and intracellular calcium signaling are essential for BCR-induced apoptosis. Because DT40 B cells are highly proliferative and readily integrate exogenous DNA by homologous recombination, gene targeting is conveniently employed to inactivate genes of interest. This chapter describes procedures for (1) generation of knockout DT40 cell line, (2) analysis of surface BCR expression, (3) detection of tyrosine-phosphorylated proteins, (4) detection of intracellular calcium mobilization, and (5) reconstitution of depleted gene by transfection.

本文言語英語
ページ(範囲)261-270
ページ数10
ジャーナルMethods in molecular biology (Clifton, N.J.)
271
DOI
出版ステータス出版済み - 2004

All Science Journal Classification (ASJC) codes

  • 分子生物学
  • 遺伝学

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