Journal of Immunological Methods
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Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays.
Luminex bead array assays are widely used for rapid biomarker quantification due to the ability to measure up to 100 unique analytes in a single well of a 96-well plate. There has been, however, no comprehensive analysis of variables impacting assay performance, nor development of a standardized pro...
Toward development of a comprehensive external quality assurance program for polyfunctional intracellular cytokine staining assays.
We report the results of External Proficiency 3 (EP3) where participating sites performed a 7-color ICS assay. On average, sites perform well in the Flow Cytometry EQAP (median score is "Good"). The most common technical issues identified by the program involve protocol adherence and data analysis;...
Stromal cell-free conditions favorable for human B lymphopoiesis in culture.
We recently found that human CD10+ lymphocytes were produced when CD34+ hematopoietic stem and progenitor cells were cultured in contact with human mesenchymal stem cells (hMSC). Further investigation revealed that it occurred even when progenitors were separated from hMSC by membrane filters. Exper...
Two-photon laser scanning microscopy imaging of intact spinal cord and cerebral cortex reveals requirement for CXCR6 and neuroinflammation i...
We introduce a novel, facile and inexpensive method for two-photon laser scanning microscopy of the intact spinal cord in the mouse by taking advantage of the naturally accessible intervertebral space. These are powerful methods when combined with gene-targeted mice in which endogenous immune cells...
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...