Abstract：
:Dispase-treated murine hybridoma cells (SP2/0-Ag14) were transfected with the G418 resistance gene bearing plasmid pSV2-neo by electropermeabilization with a high degree of efficiency. The cells were subjected to intermittent multiple high-voltage short duration (5 microsecond) DC pulses at intervals of 1 min in a weakly conducting medium followed by selection in G418-containing medium. The transfection medium, temperature, pulse duration, and voltage were empirically determined by preliminary electropermeabilization experiments. Increasing the number of pulses resulted in a higher percentage of transfected cells, but a decrease in the number of viable cells, with the optimal transfectant yield resulting when five pulses of 10 kV/cm were administered. This method allows the rapid and efficient injection of DNA into mammalian cells, and permits the rapid production of stable, drug resistant hybridoma cell lines for use in subsequent fusion experiments.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Stopper H,Zimmermann U,Neil GAdoi
10.1016/0022-1759(88)90452-8subject
Has Abstractpub_date
1988-04-22 00:00:00pages
145-51issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(88)90452-8journal_volume
109pub_type
杂志文章abstract：:An enzyme-linked immunosorbent assay (ELISA) for the quantitation of rat erythrocyte membrane sialoglycoproteins (glycophorins) has been developed. Samples of erythrocytes and reticulocytes were analysed using this assay, and response compared among them and purified glycophorins samples. A broadly homologous behaviou...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90387-m
更新日期：1993-06-18 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：:A novel latex agglutination test employing coloured latex particles is described. The latex reagent contains two or more differently coloured latex suspensions, each colour having been coated before mixing with at least one specific antibody, so that two or more antigens can be detected and identified with a single re...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90454-6
更新日期：1987-03-12 00:00:00
abstract：:A method is described to quantitate complement fragment C4ai in human plasma, synovial fluid, and urine. Samples are first precipitated with 50% saturated (NH4)2SO4 to remove cross-reactive macromolecules C4 and pro-C4. Whereas greater than 97% of C4 is removed by this precipitation step, 88% of C4ai remains in soluti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90257-x
更新日期：1981-01-01 00:00:00
abstract：:Three commercial second generation enzyme immunoassays (EIA) and a second generation immunoblot assay (RIBA-II) for detecting antibodies to hepatitis C virus (HCV) were evaluated in a study of confirmatory testing using sera referred by six Regional Blood Transfusion Centres (RTC). Of a total of 490 samples, 203 were ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,多中心研究
doi：10.1016/0022-1759(93)90182-7
更新日期：1993-04-02 00:00:00
abstract：:Antibody phage display technology is a well established method for selecting specific antibodies against desired targets. Although phage display is the most widely used method of generating synthetic antibodies, it is laborious to perform multiple selections with different antigens simultaneously using conventional ma...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.09.034
更新日期：2011-03-31 00:00:00
abstract：:A polymorphism at position 559 in the cDNA of the FCGR3A gene encoding the FcgammaRIIIa generates two allotypes with either a valine (Val) or a phenylalanine (Phe) at amino acid position 158. This polymorphism is of major importance in immunopathology and in pharmacogenetics, especially monoclonal antibody treatments....
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00123-6
更新日期：2003-06-01 00:00:00
abstract：:Multivalent antibody fragments (scFv dimers, trimers and tetramers) provide high avidity and ideal pharmacokinetics for tumour targeting applications. This protocol describes our optimised protocol for high-level bacterial synthesis of soluble antibody scFv fragments, as either monomers or multimers, using the heat-in...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00201-5
更新日期：2000-08-28 00:00:00
abstract：:Levels of acute phase cytokines secreted ex vivo by peripheral blood mononuclear cells (PBMCs) have been shown to be associated with clinical conditions or histologic lesions in renal transplant recipients. One of the limiting factors for the potential use of this assay as a diagnostic tool is the incubation time need...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.01.003
更新日期：2011-03-07 00:00:00
abstract：:An international collaborative study was conducted at ten sites to examine the performance of enzyme immunoassays (EIAs) for the quantitation of IgG1, IgG2, IgG3, IgG4 and total IgG anti-Haemophilus influenzae type b (Hib) capsular polysaccharide in human serum. All groups used the same reagents: microtiter plates coa...
journal_title：Journal of immunological methods
pub_type： 临床试验,杂志文章,多中心研究
doi：10.1016/0022-1759(92)90163-n
更新日期：1992-04-08 00:00:00
abstract：:A 24 h, highly sensitive assay for detection of interleukin 1 (IL-1) is described. A thioguanine-resistant mutant of the murine lymphoma cell line LBRM 33 was selected (LBRM TG6). When this cell line was incubated with low concentrations of PHA and IL-1 it produced interleukin 2 (IL-2). IL-2-dependent HT2 cells were c...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90389-8
更新日期：1985-05-10 00:00:00
abstract：:The potential value of fluorescein-conjugated human alpha 2-macroglobulin as a probe for studying macrophages in murine peritoneal exudate and spleen cell suspensions has been investigated using a fluorescence-activated cell sorter. These studies revealed that the number of alpha 2-macroglobulin-positive cells in the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90360-6
更新日期：1985-10-10 00:00:00
abstract：:Information about the IgE receptor and IgE content of mast cells under different conditions in vivo is essential for further understanding the functions of the mast cell-IgE system. A cytofluorometric method for measuring cell-bound IgE on individual mast cells was therefore explored using peritoneal mast cells of Spr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90151-1
更新日期：1994-12-28 00:00:00
abstract：:Progress in elucidating the pathogenesis of Helicobacter pylori gastric infection and in developing an H. pylori vaccine will be aided by an animal model in which H. pylori can be reliably detected. To validate the use of the mouse model of H. pylori infection, we determined the susceptibility of three inbred strains ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00215-5
更新日期：2000-08-28 00:00:00
abstract：:The guinea pig has a storied history as a model in the study of infectious disease and immunology. Because of reproducibility of data and availability of various reagents, inbred mice have since supplanted the guinea pig as the animal model-of-choice in these fields. However, several clinically-significant microorgani...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.01.005
更新日期：2013-03-29 00:00:00
abstract：:The sensitivity of the conventional EA (Fc) rosetting assay for detecting Fc receptors could be significantly increased by coating sheep erythrocytes (SRBC) with high concentrations of a preparation of homogeneous monoclonal IgG2b anti-SRBC antibodies. This was made possible by virtue of the fact that the monoclonal a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90218-5
更新日期：1980-01-01 00:00:00
abstract：:Cell-based assays to monitor antigen-specific T-cell responses are characterized by their high complexity and should be conducted under controlled conditions to lower multiple possible sources of assay variation. However, the lack of standard reagents makes it difficult to directly compare results generated in one lab...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.04.007
更新日期：2018-07-01 00:00:00
abstract：:An indirect rosette assay which utilizes erythrocytes coupled with purified antiimmunoglobulin (Ig) antibodies was modified into a microassay for detecting HLA allo-and xenoantibodies. The test, which is performed in microtiter plates, is specific, reproducible and can handle large numbers of samples. As an inhibition...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90059-0
更新日期：1979-01-01 00:00:00
abstract：:An enzyme-linked immunosorbent assay (ELISA) was developed for screening production of monoclonal antibodies with specificity for HBsAg. Mouse hybridoma IgG were firstly extracted from assay medium with goat anti-mouse IgG adsorbed on polystyrene beads. The specific antibody was revealed by saturation with HBs antigen...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90361-1
更新日期：1982-01-01 00:00:00
abstract：:A generally applicable method was developed for the rapid and quantitative detection of both toxin- and virus neutralizing antibodies. The method was optimized for three different biological agents, i.e., Shigella toxin, influenza viruses (A/Beying, A/Taiwan and B/Yamagata) and Chikungunya virus. The in vitro micro-cy...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90356-c
更新日期：1993-12-03 00:00:00
abstract：:Ammonium-chloride-containing solutions (AC) are routinely used to lyse red blood cells during preparation of PBMC. Although exposure to AC has been described to affect the ultrastructural appearance of large granular lymphocytes and to temporarily inhibit cytolytic activity of PBMC preparations, the cellular basis of ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00326-x
更新日期：2001-06-01 00:00:00
abstract：:Chemokines, a large family of inflammatory cytokines, have been shown to play a critical role in the regulation of angiogenesis during several pathophysiologic processes, such as tumor growth, wound healing and ischemia. Semiquantitative or quantitative angiogenesis assays are commonly utilized to screen the angiogeni...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(02)00420-9
更新日期：2003-02-01 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：:The characterization of lymphoid subsets isolated from different anatomical sites is of great importance for understanding the mechanisms and interactions of normal and pathological immune reactions in the pig. The objective of this study was to standardize a protocol for the isolation of lymphocytes from mucosal tiss...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00209-x
更新日期：2000-07-31 00:00:00
abstract：:We describe a simple and efficient non-chromatographic method for the purification of murine IgG3 and IgM monoclonal antibodies (MAbs) which takes advantage of their euglobulin properties. Following filtration, ascitic fluid is dialysed against demineralized water and centrifuged at 22,000 X g for 30 min. The resultin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90054-3
更新日期：1988-06-28 00:00:00
abstract：:The neutrophilic granulocyte is the most numerous leukocyte in peripheral blood. The development from a multipotent progenitor cell to a mature neutrophil takes place in the bone marrow over a period of 10-14 days. In order to understand the cellular mechanisms behind this process, it is necessary to investigate cells...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(99)00176-3
更新日期：1999-12-17 00:00:00
abstract：:Detection of anti-drug antibodies (ADA) can be difficult, if not impossible, in the presence of drug in the sample. This is a particular concern with therapeutic monoclonal antibodies (mAbs), which have typically longer half-lives than other proteins. For detection of ADA in presence of high drug concentrations, assay...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.01.010
更新日期：2008-05-20 00:00:00
abstract：:An evaluation of the delayed fluorescence immunoassay (Delfia) against an ELISA method for determination of diphtheria antitoxin levels in serum was performed. The Delfia was also validated in the in vivo toxin neutralisation test (Txn) in rabbits. Two variants of the Delfia were studied, a single-antigen Delfia (sDel...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00129-5
更新日期：1999-11-19 00:00:00
abstract：:The clinical value of assessing immunoglobulin (Ig)G and IgA subclasses in addition to the isotypes of soluble Igs in serum has been well established. >20years ago, the International Union of Immunological Societies and the World Health Organization performed collaborative studies in order to validate antibody (Ab) cl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.09.008
更新日期：2019-12-01 00:00:00
abstract：:Pig peripheral blood lymphocytes, typed for LD antigens by negative reactions to homozygous cells in primary MLRs and for SD antigens by complement-mediated lymphocytotoxicity, were sensitised in vitro against X-irradiated lymphocytes. These "primed" cells were rechallenged with lymphocytes possessing a range of LD an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90151-5
更新日期：1978-01-01 00:00:00
