Abstract：
:A highly sensitive fluorimetric enzyme immunoassay was developed for prostaglandin H (PGH) synthase, an intrinsic membrane protein, using n-octyl beta-D-glucopyranoside for solubilization of the synthase from the endoplasmic reticulum membrane. An anti-PGH synthase IgG-coated polystyrene ball was reacted with a PGH synthase standard or crude detergent extract of cells, washed to remove the detergent, and then incubated with anti-PGH synthase Fab'-beta-D-galactosidase conjugate. The bound beta-D-galactosidase activity was quantitated with 4-methylumbelliferyl-beta-D-galactoside, a fluorogenic substrate. The detection limit for the PGH synthase standard was found to be 1.0 pg/assay. The sensitivity of this assay is increased by about 2-3 orders of magnitude over those of previously reported radioimmunoassay or colorimetric enzyme immunoassay. The high sensitivity of the fluorimetric enzyme immunoassay allowed reliable detection of low levels of human PGH synthase present in small samples of cultured cells. Our studies with this fluorimetric immunoassay for the synthase developed a general method for determination of membrane-bound proteins at ultrasensitive levels.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Ruan KH,Kulmacz RJ,Wilson A,Wu KKdoi
10.1016/0022-1759(93)90403-tsubject
Has Abstractpub_date
1993-06-04 00:00:00pages
23-30issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(93)90403-Tjournal_volume
162pub_type
杂志文章abstract：:The reverse haemolytic plaque assay has been adapted to detect and measure the release of such cytokines as interleukin-1, -2 and -6, GM colony-stimulating factor or interferon-gamma by individual human cells derived from either peripheral blood or enzymatically dispersed breast carcinomas. Since each of these peptide...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90340-2
更新日期：1990-02-20 00:00:00
abstract：:The amino acid glutamine is an essential nutrient for cells in culture. In aqueous solutions such as liquid culture media, glutamine spontaneously decomposes into ammonia. In this study, we examined the toxicity of ammonia for two different cell lines. In mouse hybridoma cell cultures, viable cell counts were reduced ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90331-z
更新日期：1993-11-05 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：:The aim of this study was to develop an in vitro test system for pyrogenic substances. Three clones derived from human monocytoid cell lines, which were selected by their high sensitivity to lipopolysaccharide (LPS), were assessed for tumor necrosis factor (TNF) production. Their response to pyrogen-containing samples...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00112-9
更新日期：1997-09-24 00:00:00
abstract：:We performed nasal lavage (NAL) combined with induced sputum to determine exudative inflammation in the upper and lower airways in patients with chronic sinusitis and in controls. To monitor plasma exudation into the respiratory lumen and loss of size-selectivity of the mucosa, we determined the sample-to-serum ratio ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00439-2
更新日期：2001-10-01 00:00:00
abstract：:Based on the earlier observation that DNP-HSA interacts directly with C1q, a subcomponent of the first component of complement (Loos and König, 1977), evidence is presented that TNP bound to erythrocytes (E-TNP) can interact with the whole complement sequence leading to the lysis of the TNP-carrying erythrocytes; in t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90256-9
更新日期：1978-01-01 00:00:00
abstract：:A monoclonal antibody to human IgG was tested with myeloma proteins of the four IgG subclasses. When tested by immunofluorometric assay, enzyme-linked immunosorbent assay, hemagglutination and hemagglutination inhibition assays, the antibody reacted with IgG3 but not with the other three IgG subclasses. When tested by...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90317-6
更新日期：1987-02-11 00:00:00
abstract：:We report a simple procedure for the maintenance and retrieval of rodent peritoneal macrophages on dialysis membrane supports. Culture units are easily constructed from plastic petri dishes and dialysis membranes. Macrophages adhere readily but are detached easily from the dialysis membrane by mild mechanical agitatio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90322-2
更新日期：1982-11-12 00:00:00
abstract：:A unique method for assessing the in vivo recovery of cellular immunity following sublethal irradiation and syngeneic bone marrow transplantation of CBA/J mice is described. This method employs the serial surgical implantation of a urethane sponge matrix followed by impregnation of the sponge matrix with alloantigen. ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90283-2
更新日期：1985-12-17 00:00:00
abstract：:Rh0(D) antibodies which retain immune specificity after radiolabeling were prepared by a procedure which does not require IgG isolation from serum, requires 10-fold less isotope than conventional techniques and yields antibody solutions of defined composition. The method involves radioiodination of IgG on immobilized ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90226-5
更新日期：1982-01-01 00:00:00
abstract：:To provide a more defined and safer replacement for the human rabies immune globulin (HRIG) from pooled serum which is currently used for treatment of exposure to rabies virus we have developed a series of human rabies virus-specific monoclonal antibodies. Mouse-human heterohybrid myeloma cells producing rabies virus-...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00223-9
更新日期：2000-02-21 00:00:00
abstract：:An inhibitor to von Willebrand factor (vWf) was detected in the plasma from two patients with histories of mild bleeding and one patient with a severe deficiency in the Factor VIII complex using a competitive enzyme-linked immunosorbent assay (ELISA) procedure. IgG antibodies from the patients' plasmas were shown to b...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00209-8
更新日期：1997-01-15 00:00:00
abstract：:Chinese hamster lymphocyte cultures were supplemented with foetal calf serum (FCS) or hose serum (HS). Addition of 2-mercaptoethanol (2-ME) to cultures supplemented with these sera resulted in a marked increase lymphocyte proliferation. The PHA concentrations necessary for optimal lymphocyte stimulation were much lowe...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90299-x
更新日期：1982-01-01 00:00:00
abstract：OBJECTIVE:The objective of this study was the identification of the stain HIF-alpha using the Image Cytometry, and to help to count the positive cells (with HIF-alpha) and the negative cells (without HIF-alpha) from the same sample. METHOD:17 images of renal tissues from male rats of Winstar lineage; overall, there we...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.02.005
更新日期：2017-05-01 00:00:00
abstract：:A microplate enzyme-linked immunosorbent assay (microELISA) for the study of immunochemical relationships between rabbit myosin light chains is described. Purified individual fast-muscle myosin light chains (LC1F, LC2F and LC3F) and their respective antisera, obtained in chicken, were used. Optimal conditions for anti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90289-8
更新日期：1979-01-01 00:00:00
abstract：:Fungi represent emerging infectious threats to human populations worldwide. Mice and other laboratory animals have proved invaluable in modeling clinical syndromes associated with superficial and life-threatening invasive mycoses. This review outlines salient features of common vertebrate animal model systems to study...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/j.jim.2014.03.022
更新日期：2014-08-01 00:00:00
abstract：:A number of supravital fluorochromes are available to study leucocyte functions in vitro and in vivo. The fluorescein ester most widely used, fluorescein diacetate, has the disadvantage of rapid cellular efflux, whereas more recently developed fluorescent probes do not exhibit this inconvenient trait. However, their e...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90384-0
更新日期：1994-06-03 00:00:00
abstract：:We describe a simple, rapid and reproducible in vitro culture system in which human peripheral blood lymphocytes (PBLs), donated 24 h prior to initiation of culture can be stimulated to produce antigen-specific antibodies. Peripheral blood lymphocytes purified by Ficoll-Hypaque centrifugation were passed over a G10 Se...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00256-1
更新日期：1996-03-28 00:00:00
abstract：:Antisera used in immunological assay systems for small molecular weight substances are routinely prepared by coupling the hapten to a carrier protein via a chemical linker. Often this bridge is partly recognized by the antibody, resulting in reduced sensitivity when an identically structured tracer (e.g. iodine-labell...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90296-5
更新日期：1984-11-30 00:00:00
abstract：:In the course of studying the susceptibility of mice to Salmonella infection, we discovered that C3H/HeJ mice differ from other strains in both their susceptibility to infection and their response to prior vaccination. Others have noted the absence of the usual B cell stimulation by LPS in these mice. To investigate t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90108-9
更新日期：1977-01-01 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：:Flow cytometry has been utilized to evaluate the stability of antibody production by unstable subclones of a human x human x mouse heterohybridoma. Heterogeneity of cell-associated immunoglobulin heavy chain expression was demonstrated in different subclones and an increased frequency of cells containing low levels of...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90247-x
更新日期：1994-03-29 00:00:00
abstract：:alpha-Lactalbumin has been purified from mouse milk by the use of ammonium sulfate precipitation, Bio-Gel P-100 chromatography and DEAE-cellulose chromatography. Mouse alpha-lactalbumin exists as two major charge forms with the same molecular weight. Both charge forms have been used in the development of a radioimmuno...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90126-x
更新日期：1980-01-01 00:00:00
abstract：:Mixtures of sheep erythrocytes and immune spleen cells from mice were incubated in shallow slide chambers coated with erythrocyte ghosts or anti-mouse Ig, with poly L-lysine as a coupling agent. Antigen-binding cells and erythrocytes surrounding antibody-releasing lymphocytes became bound to the reactive surfaces by i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90180-1
更新日期：1978-01-01 00:00:00
abstract：:This study examined the secretion of IL-1 alpha and IL-1 beta by THP-1 leukemia cells following activation with mezerein and promotion of synthesis by interferon (IFN-gamma). Interleukin-1 (IL-1) was not detected by co-mitogenic thymocyte assays of crude supernates. Isoelectrofocusing of concentrated medium showed tha...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90266-4
更新日期：1989-09-01 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：:Contamination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis samples has commonly produced artefactual bands when sensitive detection systems, such as silver staining or immunoblotting procedures, have been used. Evidence suggests that these bands originate from human skin contamination of reagents and ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90090-7
更新日期：1988-05-25 00:00:00
abstract：:The migration inhibition by soluble PPD of peripheral blood mononuclear cells showed a much better reflection of delayed hypersentitivity to tuberculin in man than the migration inhibition of buffy coat cells. The migration index (MI) with mononuclear cells in a group of tuberculin positive subjects was 0.51 +/- 0.14 ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90012-0
更新日期：1976-01-01 00:00:00
abstract：:Serum factors which interact with human peripheral blood lymphocyte Fc gamma receptors (Fc gamma Rs) may be detected in vitro by the EA rosette inhibition assay (EARIA). This assay has been used to detect circulating immune complexes and certain alloantibodies directed against cell surface antigens situated in close p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90222-s
更新日期：1993-02-03 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
