Abstract：
:The purpose of this study was to develop a flow cytometric method for the detection of adenosine deaminase (ADA) in a single cell suspension of mononuclear cells. Anti-human ADA antibody was purified by affinity chromatography on a column of Sepharose 4B to which calf ADA was covalently linked. This antibody was used for indirect immunofluorescent staining of cells fixed in 4% paraformaldehyde. The specificity of staining was proved by substitution of anti-human ADA with normal rabbit IgG and by absorption experiments. The fluorescence profile of the cells was then analyzed by flow cytometry. Two groups of cells were studied: (a) thymocytes, tonsil cells and peripheral blood mononuclear cells (PBMC), (b) ADA-positive and ADA-deficient cell lines. In each of these populations of cells a peak of specific immunofluorescence staining for the enzyme could be easily distinguished from weak background staining of control preparations. Within each group, the cell population with higher ADA activity also displayed a greater intensity of cell fluorescence. Flow cytometry provides a means for quantitation of ADA in individual mononuclear cells.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
SenGupta S,Petsche D,Gelfand EW,Chechik BEdoi
10.1016/0022-1759(85)90017-1subject
Has Abstractpub_date
1985-06-25 00:00:00pages
155-62issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(85)90017-1journal_volume
80pub_type
杂志文章abstract：:Insulin was directly coupled and also indirectly coupled by a side chain to Sepharose 4B to form immunoabsorbents for affinity chromatography of insulin antibodies. The two direct derivatives were insulin lysyl and insulin phenylalanyl sepharose. The indirect derivative, insulin carboxyhexyl sepharose, was prepared in...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90130-2
更新日期：1977-01-01 00:00:00
abstract：:A radioimmunoassay (RIA) for antibodies to the Ro/SSA particle is described using Iodogen to radiolabel the antigenic protein moiety of Ro/SSA with 125I. RIA methods utilizing either 33% saturated ammonium sulfate (NH4)2SO4 or 3.5% polyethylene glycol to separate bound from free antigen are comparable. Either method i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90240-v
更新日期：1990-07-20 00:00:00
abstract：:This paper describes a method to enhance the sensitivity of enzyme immunoassays by the use of solid tagged latex beads. Various bead sizes (0.21 and 0.81 micron) were tested using particles which had been covalently modified (or coated) with biotinylated rabbit anti-mouse immunoglobulin antibodies. Detection limits of...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90158-5
更新日期：1988-11-10 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：:A new assay designed to quantitate antibody reactivity to specific peptides using biospecific interaction analysis (BIAcore) has been developed. Peptides of various lengths (15-40 amino acids) and isoelectric points (pI = 4.5-13) were covalently linked (immobilized) to a biosensor and interacted with polyclonal human ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90225-i
更新日期：1992-02-05 00:00:00
abstract：:The open reading frame of the α-subunit (amino acids 1-211) of human muscle nicotinic acetylcholine receptor (hAChR(α211)) was inserted into eukaryotic expression vector of pPIC9K to form a recombinant plasmid. After transformation and expression, the target protein rhAChR(α211) (recombinant hAChR(α211)) was secretive...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.04.015
更新日期：2011-09-30 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
abstract：:The apoptotic process occurs asynchronically in most cell populations and its duration is variable. Therefore, the ability to continuously monitor the death process occurring in individual blood cells before, during and following apoptosis induction is crucial in the evaluation of the efficiency of pro- or anti-apopto...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00263-1
更新日期：2003-10-01 00:00:00
abstract：:Dendritic cells are a specialized but trace population of antigen presenting cells that always have been enriched by multi-step procedures over a period of 1 or more days in tissue culture. Here we describe the isolation of dendritic cells from fresh mouse spleen suspensions using the FACS and a monoclonal antibody, N...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90318-p
更新日期：1990-10-04 00:00:00
abstract：:A simple and rapid test (LIMM, short for latex immunoassay) is described for detecting Salmonella typhi endotoxin. It involves the simultaneous binding of the antigen by two types of reagent particles contained in a micro-tube: an indicator latex particle coated with a monoclonal antibody specific for the O-9 determin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90279-5
更新日期：1990-12-31 00:00:00
abstract：:A sensitive time-resolved fluoroimmunoassay of nerve growth factor (NGF) has been developed. The method is based on the unique property of the lanthanides for delayed fluorescence, which reduces substantially the endogenous fluorescence of biological substances, because the excitation of the sample and detection of th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90334-7
更新日期：1989-08-15 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 method for quantitative RT-PCR using ELISA detection was developed and applied to the quantitation of IL-1 beta mRNA in clinical samples. To compensate for the 'tube effect' of RT-PCR, a synthetic RNA, pRSET RNA, was added to sample solutions as an internal standard and co-amplified with IL-1 beta mRNA. Sense primer...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00321-m
更新日期：1995-03-27 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：: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：:The Cellular Antigen Stimulation Test (CAST), a recent innovation in the laboratory detection of allergen sensitivity, is a functional assay based on the release of sulphidoleukotrienes (LTs) from allergen-activated circulating basophils. The current study was undertaken to establish optimum determinants of this proce...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.12.014
更新日期：2004-05-01 00:00:00
abstract：:Limited data exists regarding antibody avidity for human papillomavirus (HPV). We describe development of a multiplex electrochemiluminescent avidity ELISA for four HPV types (HPV 6, 11, 16, 18) by adding a dissociating step to our established multiplex HPV VLP ELISA. Initial experiments exploring ammonium thiocyanate...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.04.004
更新日期：2017-08-01 00:00:00
abstract：:The multiple antigen peptide (MAP) system has been proposed as a novel and valuable approach for eliciting antibodies to peptides and developing synthetic vaccines. The MAP system consists of a small immunogenically inert core matrix of lysine residues with alpha- and epsilon-amino groups for anchoring multiple copies...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90033-p
更新日期：1992-12-08 00:00:00
abstract：:Removal and/or neutralization of preformed anti-pig antibodies in non-human primate blood have been shown to prevent the hyperacute rejection of transplanted pig organs. The purpose of this study was to establish a suitable in vitro method that would allow for screening and comparison of various agents and methods pot...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00175-6
更新日期：1999-01-01 00:00:00
abstract：:Ranibizumab (Lucentis®), a humanized antigen-binding fragment (Fab) monoclonal antibody that blocks VEGF-A activity, is currently approved for the treatment of several retinal degenerative diseases. The assessment of drug pharmacokinetics (PK) is essential for evaluating exposure as it relates to drug safety and effic...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.05.007
更新日期：2018-10-01 00:00:00
abstract：:A quantitative immunocytochemical method for the measurement of islet cell cytoplasmic antibodies has been developed. The method employs human or rat pancreas, a protein A-peroxidase/diaminobenzidine secondary antibody system and an independent measurement of islet total and exocrine mean integrated absorbance by scan...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(12)80049-4
更新日期：1992-04-27 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：:Nonspecific effects of reagents used to selectively isolate lymphocyte subpopulations are most often analyzed according to their influence on cell viability and proliferation. While characterizing monoclonal anti-human B cell antibodies, we detected consistent changes in B cell function between individual donors, no m...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90352-1
更新日期：1984-02-10 00:00:00
abstract：:The existing methods for phagocytosis evaluation are inadequate for assessing all the real events occurring during phagocytosis, or for continuously following the kinetics of the process. Our purpose is to establish the use of luminous bacteria as an object for phagocytosis. The bioluminescence test offers an easy and...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90443-x
更新日期：1983-11-25 00:00:00
abstract：:We describe the use of 7-amino-actinomycin D (7AAD) to measure phagocytosis and the opsonizing capacity of serum. Heat-inactivated Candida albicans was previously stained with 7AAD and incubated with resident peritoneal macrophages. The samples were analyzed by flow cytometry and phagocytic cells were identified by th...
journal_title：Journal of immunological methods
pub_type：
doi：10.1016/s0022-1759(01)00358-1
更新日期：2001-07-01 00:00:00
abstract：:Dot-immunobinding assay for anti-calmodulin (CaM), and immunoblot assay for CaM have become feasible by proper choice of a fixative, i.e., formaldehyde vapor. The protocol for these simple and convenient immunological procedures is presented. Dot-immunobinding assay could be accomplished in 3 h, and immunoblot assay i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90089-6
更新日期：1989-12-20 00:00:00
abstract：:The reactivity of monoclonal antibodies (Mabs) raised against a cyclic peptide representing a chimeric V3 loop of HIV-1 gp120 with different peptide analogues was studied with a biosensor system (BIAcore) and by ELISA. In both assays, the Mabs cross-reacted extensively with the V3 regions of different HIV-1 strains an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90316-6
更新日期：1994-12-02 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：:Hybridomas secreting monoclonal antibodies (MAbs) against African horse sickness virus (AHSV) were generated using different AHSV antigen preparations (inactivated AHSV, semi-purified virus, and a preparation of nonstructural viral proteins) in one of three different in vitro primary immunization systems: (i) the Cel-...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00144-y
更新日期：1995-10-26 00:00:00
abstract：:The reductive methylation procedure for proteins in solution has been adapted for tritium radiolabelling of surface components of macrophages, including the Fc gamma receptor. High specific radioactivity of the receptor was obtained with no loss of IgG binding activity, tested by the solid-phase radioassay. To prepare...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90390-4
更新日期：1985-05-10 00:00:00
