Abstract：
:An immunoassay is described which allows the detection of glycosphingolipid (GSL) antigens on high performance thin layer chromatograms (HPTLC). The method involves: (1) the separation of GSL on HPTLCs; (2) incubation with specific antibodies against carbohydrate structures of GSL, and (3) the detection of specifically bound antibodies with alkaline phosphatase-conjugated second antibodies and 5-bromo-4-chloro-3-indolyl-phosphate (BCIP) as substrate. Using a monoclonal rat IgG2c antibody against Forssman GSL, a BALB/c monoclonal antibody against asialo GM2, and polyclonal rabbit antibodies against asialo GM1, it was shown that as little as 3 ng GSL antigen could be detected in a procedure taking detected in a procedure taking only 4 h to perform. The assay should be useful for screening mono- and polyclonal antibodies with potential specificity for GSL antigens, for the detection and quantification of GSL-antigens in tissue extracts, and for defining the specificity of anti-GSL antibodies.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Bethke U,Müthing J,Schauder B,Conradt P,Mühlradt PFdoi
10.1016/0022-1759(86)90038-4subject
Has Abstractpub_date
1986-05-01 00:00:00pages
111-6issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(86)90038-4journal_volume
89pub_type
杂志文章abstract：:One major obstacle in antibody production is the lack of highly purified immunogen. In this study, we describe an alternative strategy to circumvent this problem. A nucleotide sequence encoding a full-length of human survivin was cloned into pAK400cb. After transforming into an E. coli Origami B strain, survivin-bioti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.07.024
更新日期：2006-12-20 00:00:00
abstract：:The techniques of flow cytometry are being increasingly used to classify lymphocytes according to their surface antigens. Accurate analyses depend on the exclusion of non-lymphocytic contaminant cells. Using a fluorescence-activated cell sorter (FACS IV), virtually complete separation of human lymphocytes from monocyt...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90389-7
更新日期：1983-11-11 00:00:00
abstract：:Fractionation of lactoperoxidase iodinated cell surface material on miniature DEAE-cellulose columns provided a rapid method for separating labeled lipid from cell surface antigens. The procedure also removed poorly solubilized aggregates yielding a labeled preparation which demonstrated stable, reproducible immunopre...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90029-1
更新日期：1977-01-01 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 spleen colony-forming assay for the measurement of idiotype-specific transplantation resistance to MOPC-315 is described. The assay is highly quantitative, sensitive, reproducible, less time consuming and distinctly superior to conventional in vivo assays which measure tumor incidence, tumor size, and/or host surviv...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90085-6
更新日期：1978-01-01 00:00:00
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 expre...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90272-x
更新日期：1989-09-01 00:00:00
abstract：:The influence of antibody valency in a displacement immunoassay was investigated by comparing the whole antibody molecule with the corresponding Fab-fragment. The displacement immunoassay for the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) takes advantage of the cross-reactivity of monoclonal anti-2,4-D antibodie...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00211-7
更新日期：1999-03-04 00:00:00
abstract：:We describe a solid-phase fluoroimmunoassay, based on competition between europium-labeled 5 MeCyd (5-methylcytidine) and sample 5MedCyd (5-methyl-2'-deoxycytidine) for polyclonal anti-5MedCyd antibodies (rabbit). Europium labeling of antigen was performed using a novel polylysine-5MeCyd conjugate. Standard and sample...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90115-4
更新日期：1989-02-08 00:00:00
abstract：:In this study, we have developed a simple and efficient technique for the isolation of viable lymphocytes from the epithelium and stroma of small pre-neoplastic squamous intraepithelial lesions (SIL) of the uterine cervix. Following the separation of the epithelium from the stroma using dispase II, both biopsy fragmen...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00205-1
更新日期：1999-02-01 00:00:00
abstract：:A rapid method for the simultaneous measurement of neutrophil superoxide generation and beta-glucuronidase release is described. Assay of beta-glucuronidase using a fluorescent substrate is shown to be valid in the presence of reduced or unreduced ferricytochrome C, a prerequisite for the simultaneous assessment of th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90191-8
更新日期：1987-06-26 00:00:00
abstract：INTRODUCTION:Basophils play a major physio-pathological role in hypersensitivity related diseases. Basophils express high affinity Immunoglobulin (Ig) E receptors (FcεRI), IgG and complement regulatory. Basophils also have immunoregulatory activity through interaction with T cells. The aim of this study was to look for...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2020.112951
更新日期：2021-01-22 00:00:00
abstract：:A method for establishing and maintaining primary cultures of human spleen macrophages is described. The cultured cells have been characterized as macrophages by virtue of their adherence, morphology, phagocyte activity, surface Fc receptor, and the production of lysozyme. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90075-2
更新日期：1976-01-01 00:00:00
abstract：:After centrifugation of heparinised whole blood on Hypaque-Ficoll medium (s.g. 1.114, Ferrante and Thong, 1980) the specific gravities were measured of the layers of medium above and below the band of polymorphonuclear leukocytes (PMN), as well as of the PMN and sedimented erythrocytes. The erythrocytes and PMN were a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90363-2
更新日期：1987-01-26 00:00:00
abstract：:A competitive enzyme immunoassay with labelled antibodies has been developed for methotrexate (MTX). Methotrexate in the sample and a constant quantity of this hapten physically absorbed to polystyrene spheres through a methylated bovine albumin carrier were allowed to compete for a limiting amount of peroxidase label...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90353-8
更新日期：1983-05-27 00:00:00
abstract：:The possibility that immortalisation of in vitro immunised splenocytes may result in hybridomas secreting polyreactive antibodies was investigated. A panel of nine murine hybridomas, secreting IgM(kappa) anti-goat immunoglobulin G (anti-GIgG), was produced by immortalising splenocytes that had been immunised in vitro ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00481-1
更新日期：2001-12-01 00:00:00
abstract：:Dendritic Cell (DC)-based vaccination approaches in man require a reproducible DC generation method that can be performed in conformity with GMP (Good Manufacturing Practice) guidelines and that circumvents the need for multiple blood drawings to generate DC. To this end we modified our previously described method to ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(98)00208-7
更新日期：1999-02-01 00:00:00
abstract：:When five cytotoxicity methods compared the toxicity of eosinophil granule major basic protein (MBP) and melittin to K562 and HL-60 cells, strikingly discrepant results were noted. Trypan blue staining, propidium iodide/CellTrackerGreen staining and incorporation of 14C-leucine assays indicated MBP damages > 75% of ce...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00055-1
更新日期：1999-07-30 00:00:00
abstract：:A method is described for immunizing mice with a protein-cellulose complex obtained by covalent coupling of antigenic protein molecules to suspended cellulose particles. Primary immunization of the animals with the complex led to a pronounced immune response persisting for 20-30 days. Subsequent administration of the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90526-0
更新日期：1986-03-13 00:00:00
abstract：:Hybridoma cells producing monoclonal antibodies against tumor-shed antigens were generated by fusing mouse myeloma cells with spleen cells from immunosuppressed mice bearing human melanoma xenografts. Thirty-eight fusion experiments were performed at different stages of tumor growth. Hybridomas producing anti-melanoma...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90074-1
更新日期：1983-02-25 00:00:00
abstract：:The aim of this study was to establish a cytokine-free, serum-free system which would enable the long-term survival and proliferation of human peripheral blood monocytes. Monocytes were isolated from peripheral blood mononuclear cells (PBMC) by adherence to untreated plastic petri dishes and maintained up to 6 weeks i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90323-l
更新日期：1992-08-30 00:00:00
abstract：:Natural Killer cells are cells of the innate immune system that are important for the recognition and clearance of virally infected cells or tumors. Examination of the development and signaling of these cells has been severely hampered due to an inability to over-express proteins in these cells. We developed a novel t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.10.014
更新日期：2009-01-30 00:00:00
abstract：:A method is described for the simultaneous in situ detection of surface receptors and cytoplasmic granules in mast cells of frozen sections of mouse skin. Surface IgE receptors are detected after saturation of the receptors with a murine monoclonal antibody of IgE isotype. The latter is subsequently detected by monosp...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90121-9
更新日期：1987-05-20 00:00:00
abstract：:A new and simple method for quantitation of IgM of low molecular weight in sera by radial immunodiffusion in 7% agar gel is described. The test is easy to perform and permits exact determination of low molecular weight IgM concentration down to very small quantities (1 mg%). Results obtained with the method described ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90094-7
更新日期：1978-01-01 00:00:00
abstract：:A polyethylene glycol (PEG) radioimmunoprecipitation assay for human IgG is described that is sufficiently sensitive to detect 0.5 ng of IgG. This model antibody-antigen system was also used to study the stoichiometries of PEG-precipitated complexes. Our results suggest that the presence of PEG may affect the stoichio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90091-7
更新日期：1979-01-01 00:00:00
abstract：:A simple and highly sensitive analytical isoelectric focusing (IEF) technique using immunofixation with anti-C2 serum followed by silver staining has been developed in order to study simultaneously the structural polymorphism of both native C2 and C2 activation fragments (C2a MW 74,000 and C2b MW 34,000). Structural C...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90112-9
更新日期：1989-02-08 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：:This study was designed to define the conditions for expansion of functional T lymphocytes from human immunodeficiency virus (HIV)-infected subjects, with the ultimate goal of using these cells for immunotherapy. The most appropriate culture conditions for good T cell proliferation included stimulation with anti-CD3 a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00018-2
更新日期：2002-04-01 00:00:00
abstract：:The complement component C3 and the cleavage products of C3b/iC3b, C3c and C3d are used as biomarkers in clinical diagnostics. Currently, no specific antibodies are able to differentiate C3d from other fragments, although such a distinction could be very valuable considering that they may reflect different pathophysio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.02.002
更新日期：2017-05-01 00:00:00
abstract：:The influence of human hepatocyte growth factor (HGF) on the transforming growth factor beta (TGF-beta) bioassay CCL-64 was examined. HGF induced proliferation of the CCL-64 cells and potently counteracted TGF-beta-induced growth inhibition. HGF was not inactivated by transient acidification to pH 2, a commonly used p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00228-6
更新日期：1996-01-16 00:00:00
abstract：:Interactions between mouse and rat monoclonal antibodies (MAbs) in EIA have been detected using panels of mouse and rat MAbs of various isotypes. Mouse and rat antibodies of the IgG2a subclass were found to bind to each other most strongly. Immobilised IgG1 antibodies of the two species showed much less interaction. R...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90300-k
更新日期：1990-06-12 00:00:00
