E2A activity is induced during B-cell activation to promote immunoglobulin class switch recombination.
AuthorsQuong MW, Harris DP, Swain SL, Murre C
JournalEMBO J
PubMed ID10562543
'The basic helix-loop-helix protein, E2A, is required for proper early B lymphopoiesis. Specifically, in E2A-deficient mice, B-cell development is blocked at the progenitor stage prior to the onset of immunoglobulin (Ig) V(D)J recombination. Here, we demonstrate that E2A plays an additional role during peripheral B lymphopoiesis. Upon activation of primary ... More
FRET analysis of protein conformational change through position-specific incorporation of fluorescent amino acids.
AuthorsKajihara D, Abe R, Iijima I, Komiyama C, Sisido M, Hohsaka T
JournalNat Methods
PubMed ID17060916
'We designed and synthesized new, fluorescent, non-natural amino acids that emit fluorescence of wavelengths longer than 500 nm and are accepted by an Escherichia coli cell-free translation system. We synthesized p-aminophenylalanine derivatives linked with BODIPY fluorophores at the p-amino group and introduced them into streptavidin using the four-base codon CGGG ... More
Flow cytometry-based biosensor for detection of multivalent proteins.
AuthorsSong X, Shi J, Swanson B
JournalAnal Biochem
PubMed ID10933853
'Microsphere-based flow cytometric detection of cholera toxin (CT) through distance-dependent fluorescence resonant energy transfer (FRET) has been developed. Simultaneous double-fluorescence changes induced by multivalent interactions between CT and fluorophore (both fluorescence donor and acceptor)-labeled ganglioside GM1 on a biomimetic membrane surface (supported bilayers of phospholipids) can be measured by a ... More
ER-Tracker dye and BODIPY-brefeldin A differentiate the endoplasmic reticulum and golgi bodies from the tubular-vacuole system in living hyphae of Pisolithus tinctorius.
AuthorsCole L, Davies D, Hyde GJ, Ashford AE
JournalJ Microsc
PubMed ID10692127
Two fluorochromes, ER-TrackerTM Blue-White DPX dye and the fluorescent brefeldin A (BFA) derivative, BODIPY-BFA, label the endoplasmic reticulum (ER) in hyphal tips of Pisolithus tinctorius and allow its differentiation from the tubular-vacuole system at the light microscope level in living cells. The ER-Tracker dye labels a reticulate network similar in ... More
Ultrasensitive hybridization analysis using fluorescence correlation spectroscopy.
AuthorsKinjo M, Rigler R
JournalNucleic Acids Res
PubMed ID7784185
The hybridization of fluorescently tagged 18mer deoxyribonucleotides with complementary DNA templates was analysed by fluorescence correlation spectroscopy (FCS) in a droplet under an epi-illuminated fluorescence microscope at the level of single molecules. The interaction can be monitored by the change in the translational diffusion time of the smaller (18mer) primer ... More
The protooncogene MYC can break B cell tolerance.
AuthorsRefaeli Y, Field KA, Turner BC, Trumpp A, Bishop JM
JournalProc Natl Acad Sci U S A
PubMed ID15753301
The protooncogene MYC has been implicated in both the proliferation and programmed cell death of lymphoid cells, and in the genesis of lymphoid tumors. Here, we report that overexpression of MYC, as found in many lymphomas, can break immune tolerance. Mice that would otherwise be tolerant to a transgenic autoantigen ... More
Internalization of monomeric lipopolysaccharide occurs after transfer out of cell surface CD14.
AuthorsVasselon T, Hailman E, Thieringer R, Detmers PA
JournalJ Exp Med
PubMed ID10449522
Lipopolysaccharide (LPS) fluorescently labeled with boron dipyrromethane (BODIPY) first binds to the plasma membrane of CD14-expressing cells and is subsequently internalized. Intracellular LPS appears in small vesicles near the cell surface and later in larger, punctate structures identified as the Golgi apparatus. To determine if membrane (m)CD14 directs the movement ... More
Direct, ultrasensitive, and selective optical detection of protein toxins using multivalent interactions.
AuthorsSong X, Swanson BI
JournalAnal Chem
PubMed ID10366891
Three highly sensitive, selective, and reagent-free optical signal transduction methods for detection of polyvalent proteins have been developed by directly coupling distance-dependent fluorescence self-quenching and/or resonant-energy transfer to the protein-receptor binding events. The ganglioside GM1, as the recognition unit for cholera toxin (CT), was covalently labeled with fluorophores and then ... More
Stains, labels and detection strategies for nucleic acids assays.
AuthorsKricka LJ
JournalAnn Clin Biochem
PubMed ID11928759
Selected developments and trends in stains, labels and strategies for detecting and measuring nucleic acids (DNA, RNA) and related molecules [e.g. oligo(deoxy)nucleotides, nucleic acid fragments and polymerase chain reaction products] are surveyed based on the literature in the final decade of the 20th century (1991-2000). During this period, important families ... More