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

Tomoharu Yasuda, Tadashi Yamamoto

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)261-270
Number of pages10
JournalMethods in molecular biology (Clifton, N.J.)
Volume271
Publication statusPublished - Jan 1 2004
Externally publishedYes

Fingerprint

B-Cell Antigen Receptors
B-Lymphocytes
Cell Line
Apoptosis
Genes
Calcium Signaling
Gene Targeting
Homologous Recombination
Systems Analysis
Protein-Tyrosine Kinases
Transfection
Chickens
Calcium
Antibodies
DNA
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Analysis of B-cell signaling using DT40 B-cell line. / Yasuda, Tomoharu; Yamamoto, Tadashi.

In: Methods in molecular biology (Clifton, N.J.), Vol. 271, 01.01.2004, p. 261-270.

Research output: Contribution to journalArticle

Yasuda, Tomoharu ; Yamamoto, Tadashi. / Analysis of B-cell signaling using DT40 B-cell line. In: Methods in molecular biology (Clifton, N.J.). 2004 ; Vol. 271. pp. 261-270.
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