Abstract：
:A new adhesion assay was developed that utilizes buoyancy, rather than washing or centrifugation, to remove non-adherent cells. Biotinylated cells were added to wells containing cell monolayers or purified protein substrates. Non-adherent cells were then removed by floatation on a dense Percoll solution. The adherent cells were fixed tightly to the plate with a Percoll/glutaraldehyde fixative and quantitated by streptavidin: horseradish peroxidase chemistry. In a side-by-side comparison of buoyancy and washing assays, the buoyancy method detected B16F10 binding to purified fibronectin at a 4-fold lower fibronectin concentration and human umbilical vein endothelia cell (HUVEC) binding to laminin at a 10-fold lower laminin concentration than did washing assays. In cell to cell adhesion assays, the buoyancy method was able to detect significantly greater binding of mononuclear leukocytes and KM12-L4 colon carcinoma cells to IL-1 beta treated human umbilical vein endothelial cells (HUVEC). The binding of human promyelocytic leukemia HL60 cells to control and IL-1 beta treated HUVEC was the same (approximately 60%) with the buoyancy method, while a washing assay demonstrated 8-fold higher binding (51% vs. 6%) of HL60 on IL-1 beta treated cells. The buoyancy assay is useful for detecting weak cell to protein adhesion and may be useful for detecting cell to cell adhesion when background binding is sufficiently low.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Goodwin AE,Pauli BUdoi
10.1016/0022-1759(95)00187-6subject
Has Abstractpub_date
1995-12-01 00:00:00pages
213-9issue
2eissn
0022-1759issn
1872-7905pii
0022175995001876journal_volume
187pub_type
杂志文章abstract：:To select sequences complementary to their binding sites, two anti-streptokinase (SK) monoclonal antibodies (mAbs), A4.5 and A5.5, were used in biopanning of 15-mer and hexamer phage-displayed peptide libraries, respectively. mAb A4.5 inhibits the catalytic activity of streptokinase-plasminogen activator complex (SKPA...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00183-7
更新日期：2002-09-15 00:00:00
abstract：:ELISAs for urokinase plasminogen activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1) have shown that tumor levels of these molecules are prognostic parameters in breast cancer as well as other types of cancer. These ELISAs measure the total amount of the given component, including preforms, active, inac...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00008-2
更新日期：1997-04-11 00:00:00
abstract：:Antigen-specific lymphocytes are important in the immune response to viral infection. Peripheral blood mononuclear cells (PBMC) are traditionally used as a source of effector cells in most immunological studies. We described here the use of the bispecific monoclonal antibodies (BSMAB) anti CD3:CD8 (CD3,8) and anti CD3...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00510-0
更新日期：2003-03-01 00:00:00
abstract：:A modification of the COS cell transient expression system for isolating cDNAs encoding antibody defined molecules was used to clone the carcinoma antigen defined by monoclonal antibody CORC-6. Bacteria were transformed with cDNA prepared from colon carcinoma cells and immediately divided into aliquots containing appr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00155-4
更新日期：1995-10-26 00:00:00
abstract：BACKGROUND:The role of CD4(+) regulatory T cells (Tregs) and their subsets during HIV infection is controversial. Cryopreserved peripheral blood mononuclear cells (PBMC) are an important source for assessing number and function of Tregs. However, it is unknown if PBMC isolation and cryopreservation affect the expressio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2016.02.004
更新日期：2016-04-01 00:00:00
abstract：:A new sandwich immunoassay method for measuring human C5a-des-Arg was developed using monoclonal antibody specifically reactive with C5a-des-Arg. Monoclonal antibodies were obtained from a panel of hybridomas produced by fusion of mouse myeloma cells, P3 X 63-AG8,653, with spleen cells from a CBF1(C57BL/6 X BALB/c) mo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90159-1
更新日期：1987-08-03 00:00:00
abstract：:Phagocytosis plays a pivotal and essential role in host immune defense, both as a focal constituent of the innate immune response and a bridging element linking innate and adaptive immunity. Phagocytosis has been demonstrated to be critical in development, tissue remodeling, wound healing and resolution of inflammatio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.06.027
更新日期：2013-04-30 00:00:00
abstract：:A method for coating native, non-derivatized, polysaccharide (PS) onto nitrocellulose (NC) for identifying PS-specific antibodies has been developed. The new feature of this method is that PS molecules are vacuum filtered onto NC in their native state by devices that can accommodate NC of different sizes and shapes. P...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90032-b
更新日期：1991-03-21 00:00:00
abstract：:We have developed a high-expression system of recombinant human mannan-binding lectin (MBL) with CHO cells. Geneticin-resistant transformants harboring human MBL cDNA in the expression vector pNOW/CMV-A were screened by immunoblot analysis for secretion of recombinant MBL. Cloning and selection by both geneticin and m...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00190-2
更新日期：1999-01-01 00:00:00
abstract：:A method for the preservation of mouse peritoneal macrophages is described. Using 5% dimethylsulphoxide with either a cooling rate of 1 degree C/min or two-step cooling with 10 min interruption at -30 degrres C allows 80% survival as judged by pinocytic activity. An important finding was that the cells must be handled...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90139-9
更新日期：1977-01-01 00:00:00
abstract：:The aim of this study was to evaluate the influence on the results of the main variables of ELISA used for the detection of antiphosphatidylethanolamine antibodies (aPE). Forty sera from patients with either autoimmune disorders including antiphospholipid syndrome (APS) or the clinical features of APS only were assaye...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.04.002
更新日期：2010-06-30 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：:A method is described to identify specific antibody-forming cells and their immunoglobulin classes by a multilayer cluster formation. Cells producing specific antibodies are mixed with erythrocytes coated with the corresponding antigen in presence of rabbit anti-human immunoglobulins. Clusters are formed only if the m...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90334-8
更新日期：1980-01-01 00:00:00
abstract：:Macrophages can be separated from other cell types by their ability to readily attach and spread on glass or on plastic surfaces which are treated for optimal growth of cultured cells (tissue culture-treated plastic). To detach macrophages from these surfaces, techniques must be used which require prior preparation of...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90142-x
更新日期：1984-03-30 00:00:00
abstract：:We have developed a sandwich enzyme-linked immunosorbent assay (ELISA) for the determination of human heart type fatty acid-binding protein (H-FABP) in human plasma and urine using the combination of two distinct monoclonal antibodies (MAbs) directed against human H-FABP purified from human heart muscle. The total ass...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00248-u
更新日期：1995-01-13 00:00:00
abstract：OBJECTIVES:To report on a novel immunofluorescence pattern specifically associated with antibodies to SS-A/Ro. METHODS:A novel immunofluorescence pattern, herein designated SS-A/Ro pattern, was characterized as myriad discrete fine speckles throughout the nucleus. Eighty-six sequential samples presenting the SS-A/Ro p...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.01.006
更新日期：2013-04-30 00:00:00
abstract：:A method for the production of human gamma interferon (HuIFN-gamma), which is both simple and efficient, has been developed. The 2 co-inducers, A-23187 and mezerein, stimulate human peripheral blood lymphocytes to produce high levels of HuIFN-gamma in both stationary cultures and spinner cultures. The compound 2,2-dim...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90428-3
更新日期：1983-10-14 00:00:00
abstract：:Leucocyte procoagulant activity (PCA) has previously been advocated as a useful measurement of cell-mediated immune reactivity. The assay is, however, susceptible to inter- and intra-experimental variation. This investigation identifies several factors which influence the stability and reproducibility of the test. Maj...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90151-6
更新日期：1985-12-27 00:00:00
abstract：:We describe an ex vivo rat infection model to study protective immunity against Fasciola hepatica at the gut level. An exact number of newly excysted juveniles (NEJs) was injected into a gut segment with an intact blood supply and which was still attached to a live anaesthetized rat. NEJs that penetrated the gut wall ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00026-x
更新日期：1998-04-15 00:00:00
abstract：:Two sets of expression vectors were constructed that permitted the efficient expression of single-chain Fv fragments (scFvs) fused N-terminally to an enhanced mutant of the green fluorescent protein GFP+ or the red fluorescent protein DsRed in insect and mammalian cells. The vectors allowed rapid cloning of scFv fragm...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.12.001
更新日期：2004-02-15 00:00:00
abstract：:Genital infections with herpes simplex virus type 2 (HSV-2) are a source of considerable morbidity and are a health concern for newborns exposed to virus during vaginal delivery. Additionally, HSV-2 infection diminishes the integrity of the vaginal epithelium resulting in increased susceptibility of individuals to inf...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2016.09.004
更新日期：2016-12-01 00:00:00
abstract：:Affinity chromatography and subsequent ion-exchange chromatography of pooled anti-benzylpenicilloyl (anti-BPO) hyperimmune sera separated 5 different anti-BPO IgG fractions as described in the preceding paper. These fractions were tested for activities in passive hemagglutination (PHA), passive immune hemolysis (PIH),...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90163-1
更新日期：1978-01-01 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：:Hapten (trinitrophenyl) was coupled to antigen (ovalbumin). The haptenated antigen was bound by anti-ovalbumin antibody and binding was quantitated with 125I-labelled anti-hapten antibodies. Thus, with a single radioactive reagent antibodies against a variety of antigens can be detected, whilst the problems inherent i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90011-x
更新日期：1978-01-01 00:00:00
abstract：:An improved fixation method for antigen detection in lymphoblastoid cells is described. Herpesvirus papio nuclear antigen (HUPNA) could be stained in several transformed lymphoid cell lines by anti-complement immunofluorescence (ACIF). Antibody to HUPNA was detected in many human sera containing antibodies to Epstein-...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90162-9
更新日期：1981-01-01 00:00:00
abstract：:Human interleukin-7 (IL-7) is a cytokine that appears to be critical for early T- and B-cell development and although IL-7 is currently under investigation as a therapeutic agent in a variety of hematolymphopoietic disorders, there have been few instances of the detection or investigation of this cytokine using a biol...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00237-9
更新日期：2000-03-06 00:00:00
abstract：:Sera from five animal species were studied as complement source in the preparation of zymosan-C3 complexes used to detect human B lymphocytes. There was no significant difference in the determination of the percentage of ZC rosette forming cells due to differences in complement source. Human B lymphocytes could not di...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90195-2
更新日期：1976-01-01 00:00:00
abstract：:The pronase-enhanced release of 14C-labeled DNA was used to assay cytotoxicity of murine natural killer cells. YAC-1 lymphoma cells were labeled with [14C]thymidine and incubated with increasing numbers of spleen cells from nude mice. At harvest, nuclease-free pronase was used to digest damaged target cells. Increases...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90152-0
更新日期：1980-01-01 00:00:00
abstract：:The single-chain Fv fragment (scFv) is the most frequently used form of recombinant antibody. It is possible to establish clones specific to a certain target by displaying the scFv library on phages followed by biopanning against the target. For the construction of superior scFv libraries, the light-chain variable reg...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.07.003
更新日期：2013-10-31 00:00:00
abstract：:Design of automated image processing systems to determine migration characteristics of individual cells is not trivial. Every test sample requires separate recording and the analysis of individual cell tracks in two- or three-dimensional migration systems by time-lapse microscopy is extremely laborious. Here, we descr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00262-x
更新日期：2003-09-01 00:00:00
