Journal of Immunological Methods
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Stability of total and rubella-specific IgG in oral fluid samples: the effect of time and temperature.
Oral fluid (saliva) samples are increasingly used as a minimally invasive alternative to serum to detect antibody for viral diagnostics and population-based surveys of immunity to common infections. Postal collection of these samples is convenient, but it is uncertain how oral fluid antibody levels...
Heparin in plasma samples causes nonspecific binding to histones on Western blots.
We observed an artifactual reactivity on Western blots when heparin was used as the anticoagulant in collected blood specimens. This nonspecific interaction was found to be due to immunoglobulin aggregates that bound to cellular proteins, in particular histones. Nonspecific interaction was not obser...
Detection of and discrimination between total and free human interleukin-4 and free soluble interleukin-4 receptor by ELISA.
We have established a set of new ELISA systems for the measurement of human IL-4 in its distinct forms. To select suitable pairs of anti-IL-4 antibodies, a chequerboard interference analysis with six highly-selective human IL-4 specific monoclonal antibodies was performed. For the determination of t...
Bone marrow cellular composition in Listeria monocytogenes infected mice detected using ER-MP12 and ER-MP20 antibodies: a flow cytometric al...
We have examined whether double labelling of bone marrow with the anti-precursor monoclonal antibodies ER-MP12 and ER-MP20 could be used for differential analysis by flow cytometry, as these antibodies define six relatively homogeneous cell populations in mouse bone marrow. Following a sublethal inf...
Single laser flow cytometric detection of lymphocytes binding three antibodies labelled with fluorescein, phycoerythrin and a novel tandem f...
We combined a new fluorescent tandem conjugate (TC, R-phycoerythrin plus Texas red, available as DuoCHROME) with fluorescein (FITC)- and phycoerythrin (PE)-labelled monoclonal antibodies for the simultaneous detection of three antigens on individual lymphoid cells. Considerable amounts of fluorescen...
Isolation of rat/rat T cell hybrids using W/Fu C58(NT)D as fusion partner.
Functional rat/rat T cell hybrids were isolated for the first time by the fusion of spleen cells from rats tolerized to the hapten TNP to a HAT-sensitive rat thymoma (C58(NT)D). 11 fusions using different protocols were attempted to assess the optimal conditions for high hybridizatio...
Monoclonal anti-histamine antibody. Preparation, characterization and application to enzyme immunoassay of histamine.
We developed an enzyme immunoassay for histamine and pg amounts could be detected. The same assay was used to quantify the allergic release of histamine from guinea pig lung mast cells. Results obtained either by the present enzyme immunoassay or by a fluorometric assay were closely correlated (corr...
Sustained in vivo depletion of splenic langerin(+) CD8α(+) dendritic cells is well-tolerated by lang-DTREGFP mice.
We found that prolonged treatment with DT did not cause weight loss, or neutrophilia, as reported in some DT-based depletion models. Therefore, the in vivo depletion of murine langerin(+) CD8α(+) DCs can be maintained over time to analyse their function during the full course of an immune response....
Monoclonal antibody binding-site diversity assessment with a cell-based clustering assay.
We present a high through put assay that enables grouping of crude hybridoma supernatants without a need for antibody purification. In addition, the assay does not require recombinant protein, because it is conducted on cells that express the antigen of interest. Hence, one can use the antibody-clus...
Optimization of murine small intestine leukocyte isolation for global immune phenotype analysis.
We demonstrate that two different digestion protocols yield significant differences in detection of CD4(+) and CD8(+) T cells, NK cells, monocytes and interdigitating DC (iDC) populations, highlighting the importance and impact of cell collection protocols on assay outputs. The optimized protocol de...