Abstract：
:Enzyme immunoassay techniques are widely use to quantify various antigens and antibodies. The final step of these techniques (i.e. enzyme reaction) may be carried out in several ways (e.g. chromogenic, fluorogenic, or radioactive substrate and thermometric measurement). This paper compares the effectiveness of the chromogenic and the fluorogenic substrates in the beta-galactosidase immunoassay. Using microtitration plates (150 microliter samples) coated with anti-human IgE, and anti-human IgE labeled with E. coli beta-galactosidase, the lowest concentrations of IgE that one could detect employing either the chromogenic (o-nitrophenyl-beta-D-galactopyranoside) or the fluorogenic (4-methyl-umbelliferyl-beta-D-galactopyranoside) substrate were determined. It was found that both substrates were almost equally effective in measuring the lowest concentration of IgE (0.075-0.13 IU/ml) under the optimal conditions. But, using fluorogenic substrate and suitable apparatuses the enzyme immunoassay can be miniaturized. Thus by using decreasing volumes of reagents, progressively smaller amounts of antigen were quantified: as the sample volumes were reduced from 150 to 10 microliter and finally to 0.3 microliter a progressive decrease from 7 x 107 molecules of IgE to 2.9 x 107 molecules and to 1.5 x 106 molecules was observed. The corresponding lowest detection limits were 0.075 IU/ml, 0.46 IU/ml and 0.8 IU/ml.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Labrousse H,Guesdon JL,Ragimbeau J,Avrameas Sdoi
10.1016/0022-1759(82)90188-0subject
Has Abstractpub_date
1982-01-01 00:00:00pages
133-47issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(82)90188-0journal_volume
48pub_type
杂志文章abstract：:A cell membrane permeabilizing treatment is described which involves the use of lysolecithin at low concentration in acidic acetate buffer and paraformaldehyde fixation. It preserved well-separated scatter cytograms of small and large lymphocytes. The accuracy of the immunochemical detection of internal antigens by fl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90162-9
更新日期：1991-04-25 00:00:00
abstract：:Irradiated human peripheral blood lymphoid cells undergo apoptosis and progressively exhibit typical changes in light scatter and plasma membrane integrity that can be easily tracked by flow cytometry. Using this model, we assessed the capacity of a newly developed fluorochrome, Apostain, in identifying early apoptosi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00054-9
更新日期：1997-06-23 00:00:00
abstract：:Phagocytosis plays a pivotal and essential role in host immune defense, both as a focal constituent of the innate immune response and a bridging element linking innate and adaptive immunity. Phagocytosis has been demonstrated to be critical in development, tissue remodeling, wound healing and resolution of inflammatio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.06.027
更新日期：2013-04-30 00:00:00
abstract：:Measurement of the absorbance due to light scattering at 40 nm proved to be a simple and reliable way of assessing the extent of aggregation in heat-treated IgG solutions. Using this technique the rate of aggregation was demonstrated to be markedly temperature dependent with a sharp inflexion in the curve close to 63 ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90274-x
更新日期：1981-01-01 00:00:00
abstract：:Langerin CD207 is a type II transmembrane protein. It is responsible for the formation of Birbeck granules, which are intracellular organelles within Langerhans cells, the dendritic cells of stratified squamous epithelia like the epidermis. Because current anti-CD207 antibodies have limitations, we prepared new monocl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2007.05.001
更新日期：2007-07-31 00:00:00
abstract：:A method is described for measuring antibody-dependent cellular cytotoxicity (ADCC) of mononuclear cells from human peripheral blood using an established murine cell line and commercially prepared antisera. The test utilizes a standard 51Cr release technique. The ADCC activity of mononuclear cells obtained from 10 hea...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90107-3
更新日期：1985-05-23 00:00:00
abstract：:The monoclonal antibody-specific immobilization of lymphocyte antigens (MAILA) assay was developed to detect antibodies present in human alloantisera against antigens of different major histocompatibility complex loci, particularly of class II specificity. The MAILA assay has been used in our laboratory to the determi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90094-n
更新日期：1993-01-04 00:00:00
abstract：:The haem enzyme myeloperoxidase (MPO) (EC 1.11.1.7) with a spectral A430/A280 ratio greater than 0.7 and a specific activity of 125 U/mg was purified from isolated human neutrophils. To obtain a radioimmunoassay (RIA) for this enzyme, a specific antiserum against human neutrophil MPO was raised in rabbits and used at ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90023-9
更新日期：1991-03-21 00:00:00
abstract：:Mixtures of sheep erythrocytes and immune spleen cells from mice were incubated in shallow slide chambers coated with erythrocyte ghosts or anti-mouse Ig, with poly L-lysine as a coupling agent. Antigen-binding cells and erythrocytes surrounding antibody-releasing lymphocytes became bound to the reactive surfaces by i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90180-1
更新日期：1978-01-01 00:00:00
abstract：:Macrophages are an important part of the cellular immune system and play a key role during immune responses. Thus, macrophages are interesting targets in basic and clinical research. Primary monocytes or monocyte-derived macrophages do not proliferate on a suitable scale so that their use for functional studies in vit...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2009.03.014
更新日期：2009-05-31 00:00:00
abstract：:The indirect immunofluorescent technique has been shown to be a more sensitive method than either immunoelectrophoresis or gel filtration for testing the efficacy of the gel filtration method in fractionating human immunoglobulins. It has been confirmed that IgM fractions of some sera from patients with multiple scler...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(75)90078-2
更新日期：1975-01-01 00:00:00
abstract：:To reduce the cost, time and waste in screening for HIV and HBV infections a combined assay for HIV-1 and -2 antibodies and HBsAg has been developed. Monoclonal anti-HBs antibodies were co-immobilized with synthetic peptides representing immunodominant regions of HIV-1 and -2. The presence of anti-HIV antibodies in th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90262-6
更新日期：1993-01-14 00:00:00
abstract：:A method is described for determining the percentage of rosettes formed by sheep erythrocytes and human peripheral lymphocytes (ER) using cytofluorographic analysis (CFGA). The technique utilizes acridine orange dye for differentially staining nucleus and cytoplasm of the lymphocytes to distinguish them from erythrocy...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90091-0
更新日期：1976-01-01 00:00:00
abstract：:Many antigens are shared between different types of human leukocytes. In an effort to obtain new lineage-specific monoclonal antibodies, particularly antibodies to dendritic cells, we attempted to tolerize newborn mice to one type of leukocyte and then immunize the adults with another. We found that T cells, either un...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00335-t
更新日期：1995-04-12 00:00:00
abstract：:Mast cells are important effector cells of allergy and are involved in the pathology of many other diseases. Measurement of β-hexosaminidase activity, the most commonly used method for evaluation of murine mast cell activity, requires a large number of cells and thus is of limited utility for studying mast cells in mo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.02.014
更新日期：2012-05-31 00:00:00
abstract：:We report the development of a simple, efficient, expedient, and inexpensive procedure for isolating a large and relatively pure population of macrophages (Mphi) from residual (i.e., non-tumor) lung tissue obtained from lung cancer patients undergoing either a lobectomy or pneumonectomy. The proposed technique was fou...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90011-6
更新日期：1994-09-14 00:00:00
abstract：:In general, antigen affinity columns made with commercially prepared activated affinity supports bind antibody specific for the coupled antigen. Nonetheless, in some cases affinity purification may yield antibodies to molecules other than the molecule of interest. In this report, we demonstrate such an occurrence: an ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90246-c
更新日期：1991-01-24 00:00:00
abstract：:The methods of cell detachment influence phenotype and function of human macrophages cultured in vitro. However, comparative studies defining the influence of cell detachment techniques on secondary characterization of M1 or M2 polarized macrophages are largely absent from the literature. In this study we evaluated th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.08.001
更新日期：2015-11-01 00:00:00
abstract：:Mold contamination and exposure to fungi in indoor environments has been associated with various adverse health effects but little is known about the significance of individual fungal species in the initiation or exacerbation of such effects. Using Stachybotrys chartarum as a model fungus we sought to demonstrate that...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2005.12.001
更新日期：2006-02-20 00:00:00
abstract：:A system proposed for measurement of mutational risk consists in detection of hemoglobin mutations expressed in erythrocytes. For this detection the production of antibodies specific for Hb variants is essential. Recently we reported a sensitive solid-phase EIA for the production and selection of polyclonal and monocl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90189-8
更新日期：1983-05-13 00:00:00
abstract：:A new quantitative assay for migration inhibitory factor (MIF) employs 3H-labelled cultured mouse or human lymphoid cells migrating from capillary tubes. Capillaries filled with labelled cells are placed in liquid scintillation counting vials, along with the MIF-containing sample and are removed at the end of a five-h...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90258-2
更新日期：1978-01-01 00:00:00
abstract：:Purified polyclonal immunoglobulin preparations representing the 4 rat IgG subclasses were tested for binding to Staphylococcus aureus protein A attached to 3 different solid phases (Staphylococcus aureus Cowan I bacteria, Sepharose CL4B and Sepharose 6MB). Protein A Sepharose CL4B showed higher reactivity with IgG1 a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90251-x
更新日期：1983-08-26 00:00:00
abstract：:A microenzyme-linked immunosorbent assay (microELISA) method has been developed using an Astroscan 2100 system automated plate reader which was initially designed for tissue typing by a two colour fluorescent microcytotoxicity assay. A 96-well plate ELISA used for screening mouse monoclonal antibodies raised against s...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(12)80043-3
更新日期：1992-04-27 00:00:00
abstract：:The metaphase index assay (MIA) for thyroid growth stimulators, as originally described, used FRTL-5 thyroid cells cultured in Bellco culture chambers and glass microscope slides. The metaphases were observed using the nuclear strain aceto-lacto orcein. However the surface properties of the glass proved to be variable...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90067-1
更新日期：1988-06-28 00:00:00
abstract：:Natural killer (NK) cells are spontaneously cytotoxic immune effector cells with the ability to selectively destroy tumor cells without harming normal cells. To perform this function, NK cells utilize two main cytotoxicity pathways, the well known perforin/granzyme-mediated secretory/necrotic killing and the recently ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00361-1
更新日期：2001-07-01 00:00:00
abstract：:The technology of the mixed antiglobulin rosetting (MARR) and the direct antiglobulin rosetting reaction (DARR) is reviewed. These tests have been found to be more sensitive than the standard direct immunofluorescence (DIF) method for delineation of Ig-bearing B lymphocytes and to have practical advantages over DIF. F...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(81)90268-4
更新日期：1981-01-01 00:00:00
abstract：:Human vascular endocan is a proteoglycan exhibiting tumorigenic activity through both its glycan and protein cores. Endocan mRNA is identified as being one of the most significant molecular signatures defining a poor prognosis in lung, breast, kidney, prostate, and thyroid malignancies. The survival inversely correlat...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.02.009
更新日期：2012-04-30 00:00:00
abstract：:A fast and simple indirect immunoperoxidase staining technique is described, which can also be used for the characterization of cells with high endogenous peroxidase activity such as many haemopoietic cells. It is based on the combination of a newly developed glucose-oxidase plus glucose procedure for the inhibition o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90267-x
更新日期：1986-01-22 00:00:00
abstract：:Interferon gamma (IFNγ) is a cytokine involved in many anti-viral and immunoregulatory processes. One of the major mechanisms through which IFNγ exerts these effects is by inducing expression of indoleamine 2,3 dioxygenase-1 (IDO1), an enzyme that catalyses the first, rate-limiting step of the kynurenine pathway. In t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.05.011
更新日期：2013-08-30 00:00:00
abstract：:A mixed hemadsorption (MH) test for naturally acquired rubella immunity and immune responses after rubella vaccination has been compared with the hemagglutination-inhibition (HI) test, the immunodiffusion (ID) test, the neutralization test (NT), the hemolysis-in-gel (HIG) test and the indirect immunofluorescence (IF) ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90282-3
更新日期：1980-01-01 00:00:00
