Abstract:
:Most current methods for measuring cell death are based on plasma membrane disintegration and the consequent release of cytoplasm. The relevant cells are usually loaded with a label (usually 51Cr or 125I), the release of which is measured. I describe here a method, based on the recent evidence that dying cells often degrade their DNA into small fragments, which measures the DNA retained by living cells rather than the cellular components lost by dying cells. The assay is set up essentially like the current cell lysis assays and harvested like a cell proliferation assay. It is faster, more sensitive, easier to set up, less expensive and safer than the current standard 51Cr release assay.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Matzinger Pdoi
10.1016/0022-1759(91)90325-asubject
Has Abstractpub_date
1991-12-15 00:00:00pages
185-92issue
1-2eissn
0022-1759issn
1872-7905pii
0022-1759(91)90325-Ajournal_volume
145pub_type
杂志文章abstract::A simple method is described for the preparation of vimentin-type intermediate filaments for solid-phase immunoassays. Bovine aortic endothelial cells were grown to confluency in multiwells, and extracted with Triton X-100 and a high-salt buffer. Residual protein in the wells consisted of 53% vimentin as judged by SDS...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90149-8
更新日期:1985-12-27 00:00:00
abstract::This report describes the use of biotin-labeled antibodies in sensitive enzyme immunoassay systems for the measurement of bacterial antigens. Biotinylated immunoglobulins could be reproducibly formulated using N-hydroxysuccinimide biotin ester to link biotin to the immunoglobulin. Binding of the biotinylated antibody ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(83)80021-0
更新日期:1983-02-11 00:00:00
abstract::We describe techniques to detect nuclear transcription factors in thymocyte subsets using flow cytometry. We have adapted a method that minimizes autofluorescence of fixed cells, thereby allowing the detection of proteins expressed at low levels. An accompanying method in which the cytoplasm is removed from stained ce...
journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:2003-08-01 00:00:00
abstract::Methods for the dissociation of immunoglobulin-Brucella complexes (IgM-rich) were evaluated. The antigen-antibody complex was separated from serum by centrifugation and dissociated with either H2O, 2.0 M pH 2.0 glycine buffer, or various concentrations of KSCN. Optimal antibody recoveries for KSCN were obtained using ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90152-2
更新日期:1979-01-01 00:00:00
abstract::Cow's milk protein allergy (CMPA) is one of the most prevalent human food-borne allergies, particularly in children. Experimental animal models have become critical tools with which to perform research on new therapeutic approaches and on the molecular mechanisms involved. However, oral food allergen sensitization in ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2012.04.007
更新日期:2012-07-31 00:00:00
abstract::The use of therapeutic antibodies, delivered by intravenous (IV) instillation, is a rapidly expanding area of biomedical treatment for a variety of conditions. However, little is known about how the antibodies are anatomically distributed following infusion and the underlying mechanism mediating therapeutic antibody d...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2017.07.009
更新日期:2017-11-01 00:00:00
abstract::Accurate detection of apoptotic cells is important for the determination of cell viability. The aim of this study was to compare the sensitivity of the cell permeant SYTO 16 fluorescent dye for detecting early apoptotic mononuclear cells (MNCs) in normal donor blood with other apoptosis assays [i.e. Annexin-V, light s...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2005.07.017
更新日期:2005-10-30 00:00:00
abstract::Hereditary angioedema (HAE) is an autosomal dominant disease characterized by recurrent episodes of potentially life-threatening angioedema. The most widespread underlying genetic deficiency is a heterozygous deficiency of the serine protease inhibitor C1 esterase inhibitor (C1-Inh). In addition to low C4 levels, the ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.06.004
更新日期:2008-09-30 00:00:00
abstract::Dispersed fluorocarbon oil droplets were used as the antigen-carrier particle in an immunological agglutination assay. The extent of agglutination was found to vary with the percentage of antigen to that of inert protein used in the solution to coat and stabilize the oil droplets. Maximum sensitivity was observed when...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90533-8
更新日期:1986-03-13 00:00:00
abstract::Murine B16 melanoma cells and metastasis variants of this tumor are resistant to NK activity mediated by normal splenocytes. The B16 cells are, however, sensitive to splenocyte-mediated cytostasis. Cytostasis was measured by a [125I]UDR incorporation-inhibition [(125I]UDR I-I) assay. The main characteristics of the [1...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90066-1
更新日期:1981-01-01 00:00:00
abstract::Use of whole blood for lymphocyte proliferation experiments have advantages over the usual isolated lymphocyte procedure. We have systematically optimized methodology for whole blood lymphocyte proliferation in male Sprague-Dawley rats. Both phytohemagglutinin (PHA) and concanavalin A (ConA) stimulate lymphocyte blast...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00084-n
更新日期:1995-08-18 00:00:00
abstract::Most monoclonal antibodies to mouse antigens have been derived from rat spleen-mouse myeloma fusions. Many resultant hybridomas express one of several myeloma kappa chain transcripts, even though the parent myeloma may have been ascribed as not expressing light chain protein. Previous reports have only differentiated ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2006.07.002
更新日期:2006-08-31 00:00:00
abstract::We report a restainin method for restoring fluorescence in paraffin-embedded tissue sections previously treated with the indirect immunofluorescence technique. Antisera to gastrin and group II pepsinogens were used. Fluorescence was restored in completely faded sections retrived from storage files, as well as in secti...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90121-1
更新日期:1977-01-01 00:00:00
abstract::The identification and characterization of human mast cell (MC) functions are hindered by the shortage of MC populations suitable for investigation. Here, we present a novel technique for generating large numbers of well differentiated and functional human MCs from peripheral stem cells (=peripheral stem cell-derived ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2014.07.003
更新日期:2014-11-01 00:00:00
abstract::Laurell crossed immunoelectrophoresis (two-dimensional electroimmunodiffusion) was used to prepare minute amounts of purified parasite antigens complexed with their precipitating antibodies obtained from rabbits. These complexes, emuslified in Freund's complete adjuvant, were then used to prime rabbits for selective p...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(78)90273-9
更新日期:1978-01-01 00:00:00
abstract::Antibodies to a wide range of self-antigens, including those directed against nucleic acids or nucleic acid-binding proteins are the essential biomarkers for diseases such as systemic lupus erythematosus (SLE). Highly complex libraries of nonamers consisting of N-substituted glycines (peptoids) were screened for compo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2013.11.004
更新日期:2014-01-15 00:00:00
abstract::A method of data analysis for flow cytometry is presented which enables up to eight-dimensional data to be handled by a microcomputer with 28K addressable plus a further 32K non-addressable memory. The multi-parameter coordinates are coded into single numbers using a minimal modification of the array vector mapping eq...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90333-x
更新日期:1988-10-26 00:00:00
abstract::Regulatory T cells (Treg) have recently come to the fore in studies of immune regulation, particularly in autoimmune disease and cancer. While there appear to be several distinct subsets of T cells with regulatory function, a population described as natural Treg and characterized by expression of the transcription fac...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2007.06.006
更新日期:2007-10-31 00:00:00
abstract::There are several double immunolabelling methods but each has its drawbacks. More often than not, antibodies with the required specificities are available in only one species and their use normally produces false labels due to cross-reactivity. We describe a new and reliable technique for staining with primary antibod...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00281-2
更新日期:1996-04-19 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::The purpose of this study was to compare and contrast two enzyme immunoassay systems: the enzyme-linked immunosorbent assay (ELISA), which utilizes polystyrene microtiter plates as the adsorptive surface and the enzyme-linked immunoflow assay (ELIFA), which utilizes nitrocellulose membranes. The principal parameter un...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90152-6
更新日期:1991-08-09 00:00:00
abstract::Anti-drug neutralizing antibodies (NAbs) formed due to unwanted immunogenicity of a therapeutic protein point towards a mature immune response. NAb detection is important in interpreting the therapeutic's efficacy and safety in vivo. In vitro cell-based NAb assays provide a physiological system for NAb detection, howe...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2015.02.006
更新日期:2015-04-01 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::For microcytotoxicity, when numbers of effector or target cells are limited, the method of Takasugi and Klein (1970) is at present the most convenient way of assessing cytotoxicity. In this procedure, target cell survival is affected by minor variations in culture conditions or washing vigour and the variations may be...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90384-2
更新日期:1982-01-01 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::Assays to detect autoantibodies to glutamic acid decarboxylase (GAD65) and the protein tyrosine phosphatase-like molecule IA-2, which are both present in pancreatic islets, have been used in the diagnosis and prediction of type 1 diabetes. In this study a novel fusion protein combining the entire GAD65 molecule with t...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00303-3
更新日期:2000-12-01 00:00:00
abstract::Enzyme-linked antibody was used to treat unfixed herpesvirus-infected human fetal lung cell cultures in a mode which permitted the visualizing of local sites of infection. Foci containing as few as 20 herpesvirus-infected cells produced sufficient viral mass to be easily detectable by this method. 'Clouds' or 'plumes'...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90361-6
更新日期:1981-01-01 00:00:00
abstract::Recent data generated in our laboratory indicates that human lymphokine-activated killer (LAK) cells can be induced in several commercially available serum-free medium formulations. Interestingly, LAK cells were also generated in RPMI 1640 medium plus 1000 U/ml IL-2 without addition of human serum. However, higher lev...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90239-6
更新日期:1988-05-09 00:00:00
abstract::Thrombospondin (TSP), a platelet glycoprotein, was purified from platelet supernatants applied sequentially to Sepharose 4B and heparin-agarose affinity columns. This TSP inhibited alternative pathway activity in serum as assessed by lysis of rabbit erythrocytes and contained bound heparin, a substance which is known ...
journal_title:Journal of immunological methods
pub_type: 信件
doi:10.1016/s0022-1759(97)00006-9
更新日期:1997-03-28 00:00:00
abstract::Three different methods for the radioiodination of surface proteins in trypanosomatids have been comparatively analyzed. Data interpretation in the lactoperoxidase-mediated method was impaired by autoiodination of the enzyme and its tight retention by the cells. In the enzymobeads method, iodinated enzymes were releas...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90051-5
更新日期:1982-07-30 00:00:00