Abstract:
:Direct application of membrane fractions to a nitrocellulose support without previous solubilization, denaturation or fixation of the antigen permits the quantitation of antibodies binding to denaturation sensitive epitopes. In experiments reported here, this method was used to determine the rate of EGF receptor expression on squamous carcinoma cell lines. The assay revealed that dexamethasone treatment leads to an eight-fold increase of EGF receptor protein expression on C4-I cervical carcinoma cells. The data, when compared to the results obtained in immunoprecipitation experiments, suggest that this simple protocol yields reliable and precise quantitative data. The simplicity of the method permits simultaneous testing of large sample numbers with various antibodies.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Von Knebel Doeberitz M,Drzonek H,Koch S,Becker CMdoi
10.1016/0022-1759(89)90272-xsubject
Has Abstractpub_date
1989-09-01 00:00:00pages
259-64issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(89)90272-Xjournal_volume
122pub_type
杂志文章abstract::Radioactive double labeling was adapted for the analysis of mediators of cellular immunity. Two identical lymphocyte cultures were simultaneously labeled with [3H]- or [14C]leucine. Each of the cultures was stimulated with antigen or mitogen. The combined supernatants were then subjected to various fractionation proce...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(78)90177-1
更新日期:1978-01-01 00:00:00
abstract::During a phase III pertussis vaccine trial, serum antibody responses were measured by two enzyme-linked immunosorbent assays (ELISA) for pertussis toxin and filamentous haemagglutinin. These were used both for studies of antibody levels after vaccination and for diagnostic purposes. Since the absorbance values obtaine...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00067-k
更新日期:1995-06-28 00:00:00
abstract::A new antibody overlay technique for 2-dimensional immunoelectrophoresis (2-D-IEP) is described. After the first dimension electrophoresis of the antigen, the desired amount of antiserum is applied to the initial agarose layer and then evenly distributed over a defined surface of the gel with a PVC film. This modifica...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90332-6
更新日期:1984-02-10 00:00:00
abstract::We describe the preparation of a new rat monoclonal antibody (CRC1) to the N-terminal sequence of the 43 kDa subunit of human ovarian inhibin, and its use together with other anti-peptide monoclonal antibodies, in two-site immunoassays for the detection of inhibin-related material in biological fluids. The Fab fractio...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90342-5
更新日期:1993-10-15 00:00:00
abstract::As a model for the isolation of a labile or trace protein, the purification of uteroglobin (UGL) by immunoaffinity chromatography is described. Antibody was isolated from sheep antiserum by immunoprecipitation, and coupled to divinylsulphone-activated agarose (Mini Leak). For the immunoabsorption stage rabbit uterine ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(89)90400-6
更新日期:1989-05-12 00:00:00
abstract::A practical mixed hemadsorption assay (MHA) was developed for the detection of cell surface binding anti-tumor antigen antibodies (anti-TA Ab) in the human sera. The assay was compared to enzyme-linked immunosorbent assay (ELISA) and dot ELISA in the detection of the antibodies developed against GA733-2 antigen. SW111...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00560-9
更新日期:2002-03-01 00:00:00
abstract::Two murine IgM monoclonal antibodies (MAs) directed against the lipid A portion of bacterial lipopolysaccharide (LPS) were compared in their binding to Re LPS and lipid A and their idiotypic make-up with rabbit anti-idiotypic sera. Horseradish peroxidase (HRPO)-labelled MAs 8-2 and 26-20 bound equally well to Re LPS. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90308-i
更新日期:1990-06-12 00:00:00
abstract::A competitive RT-PCR method that permits reliable quantification of minute amounts of reverse-transcribed mouse lymph node mRNA is described. Using this technique, an absolute number of cDNA copies ranging from 10(3) to 10(5) can be determined, with a precision superior to 25%. The standard templates described in the ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00186-5
更新日期:1997-12-29 00:00:00
abstract::Quantitative indirect immunofluorescence (QIIF) methods used to measure absolute numbers of surface-expressed antigens have produced conflicting results [Marchant, A., Duchow, J., Delville, J., Goldman, M., 1992. Lipopolysaccharide induces up-regulation of CD14 molecule on monocytes in human whole blood. European Jour...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00087-3
更新日期:1999-08-31 00:00:00
abstract::A recombinant retroviral vector with an inserted bacterial neomycin resistance (neo) gene was used to transfer in vitro neomycin resistance (neoR) into murine cytotoxic lymphocyte precursors (CLP). The infection protocol involved co-cultivation of mitogen-activated splenic T-blasts with irradiated cells that produced ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90036-0
更新日期:1986-05-01 00:00:00
abstract::We have developed a sensitive and flexible method for the qualitative evaluation of IgG-Fc receptor interactions in cell suspensions. The assay is based on the flow cytometric quantitation of antibody-coated erythrocyte (EA) rosetting using fluorescein-labelled indicator erythrocytes (E). The number of IgG molecules o...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90070-c
更新日期:1990-03-09 00:00:00
abstract::The automated complement fixation system presented in this paper differs from those described by Studievic et al. (1971) and Vargues and Henley (1974). In the one developed here only one sampler is used with a wheel presenting a double row of cups. This system can assay 60 samples per h and can be modified to double t...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(76)90183-6
更新日期:1976-01-01 00:00:00
abstract::The periodate oxidation and chromium chloride coupling methods were compared for their ability to sensitize indicator erythrocytes with staphylococcal lipoteichoic acid (LTA) for the detection of specific antibody. Erythrocytes, sensitized with periodiate-activated lipoteichoic acid, were found to be superior for use ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90070-8
更新日期:1980-01-01 00:00:00
abstract::A study was undertaken to determine whether large amounts of blood (antiserum) could be rapidly removed from rabbits over a two-month period using a simple suction technique. It was found that half a blood volume could be readily removed each week (over 900 ml/rabbit/54 days) without a single fatality. The rabbits eff...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(78)90081-9
更新日期:1978-01-01 00:00:00
abstract::A luminescence immunoassay (LIA) has been developed utilizing the chemiluminescent luminol reaction with heme as catalyst. Rabbit antibody against human serum albumin was quantitated in antigen coated plastic tubes using commercially available goat anti-rabbit IgG conjugated to horseradish peroxidase which was the sou...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90048-6
更新日期:1979-01-01 00:00:00
abstract::Delineation of the immune correlates of protection in natural infection or after vaccination is a mandatory step for vaccine development. Although the most recent techniques allow a sensitive and specific detection of the cellular immune response, a consensus on the best strategy to assess their magnitude and breadth ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2011.09.005
更新日期:2012-01-31 00:00:00
abstract::The measurement of the Epstein-Barr viral load in peripheral blood has been recognised as an important way of monitoring the response to treatment in patients with Epstein-Barr virus (EBV)-related malignancies. In particular, EBV load in transplant recipients can be used as a predictive parameter for Post-transplant L...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(02)00333-2
更新日期:2002-12-15 00:00:00
abstract::We describe a highly sensitive, non-radioactive assay for T cell activation, based on the rapid induction of class II MHC expression by constitutively negative bovine endothelial cells, when cultured in the presence of supernatants derived from activated bovine T cells. We demonstrate the effectiveness of this assay i...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00187-3
更新日期:2000-05-26 00:00:00
abstract::Modifications of the classical technique for double diffusion in agar plates are presented. These procedures allow insoluble proteins to be dissolved in a variety of denaturing solvents and antibody production specificity to be monitored by the appearance of precipitin lines in agarose gels as in the conventional Ouch...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90192-2
更新日期:1977-01-01 00:00:00
abstract::The native human antibody repertoire holds unexplored potential for the development of novel monoclonal antibody therapeutics. Current techniques that fuse immortal cells and primary B-lymphocytes are sub-optimal for the routine production of hybridomas that secrete human monoclonal antibodies. We have found that a mu...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2004.05.005
更新日期:2004-08-01 00:00:00
abstract::The presence of KIR4.1 antibodies has been proposed to be a characteristic of Multiple Sclerosis (MS). This could have a significant impact on disease management. However, the validation of the initial findings has failed till date. Conflicting results have been attributed to difficulties in isolating the lower-glycos...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2017.03.008
更新日期:2017-06-01 00:00:00
abstract::A dissociation-enhanced lanthanide fluoroimmunoassay employing europium-streptavidin and time-resolved fluorimetry was developed to measure binding of biotin-labeled peptides to class II MHC proteins. Binding of biotin-peptides as measured by this assay was saturable and inhibited in the presence of unlabeled peptide....
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90124-p
更新日期:1993-08-09 00:00:00
abstract::A microELISA for the estimation of human serum thymosin alpha 1 is described. In this assay, antibody to thymosin alpha 1 is pre-incubated with the standard or serum at 4 degrees C. Unbound antibody in the liquid-phase then binds with solid-phase thymosin alpha 1. The method is sufficiently sensitive for measuring ser...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90163-2
更新日期:1985-06-12 00:00:00
abstract::We studied the binding of dog immunoglobulins G, A, M and E to protein A and protein G. Passive cutaneous anaphylaxis (PCA) testing was used for the measurement of dog IgE and enzyme-linked immunosorbent assays (ELISA) were used for the measurements of dog IgG, IgA and IgM. Protein A from lyophilized cells of Staphylo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90335-d
更新日期:1991-12-15 00:00:00
abstract::Giant liposomes (mean diameter 5.5 microns) composed of egg phosphatidylcholine or distearoyl phosphatidylcholine, phosphatidyl glycerol, cholesterol and triolein were prepared by a double emulsion technique. They were then mixed with model particulate (killed Bacillus subtilis, and killed Bacille Calmette-Guérin) and...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90368-h
更新日期:1993-12-03 00:00:00
abstract::A new, 10-fold more sensitive method, based on an improved immunofixation technique, has been devised to detect oligoclonal IgM bands in unconcentrated cerebrospinal fluid (CSF). Using agarose gel electrophoresis, 5 microliters of an unconcentrated sample containing oligoclonal bands was separated and blotted on to a ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00023-9
更新日期:1997-04-25 00:00:00
abstract::Functional analysis of antigen-specific CD8(+) T cells is important for understanding the immune response in various immunological disorders. To analyze CD8(+) T cell responses to a variety of antigens with no readily defined peptides available, we developed a system using CD4(+) phytohemagglutinin (PHA) blasts transd...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2006.05.009
更新日期:2006-07-31 00:00:00
abstract::The identification of naturally processed peptides presented by molecules of the major histocompatibility complex (MHC) has progressed significantly over the past decade. The elution of peptides from immunoaffinity purified complexes of MHC class I or class II molecules has provided highly specific biochemical informa...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/s0022-1759(00)00361-6
更新日期:2001-03-01 00:00:00
abstract::BlockLogo is a web-server application for the visualization of protein and nucleotide fragments, continuous protein sequence motifs, and discontinuous sequence motifs using calculation of block entropy from multiple sequence alignments. The user input consists of a multiple sequence alignment, selection of motif posit...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2013.08.014
更新日期:2013-12-31 00:00:00
abstract::An approach has been devised for analysing data generated in the study of in vivo cytotoxic T cell responses. The method calculates the total number of lytic units generated in the peritoneal cavity, or in cultures of the lymph nodes draining the rear footpad, following antigenic stimulation. The assessment of group d...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90387-3
更新日期:1983-11-11 00:00:00