Abstract：
:Phycobilisomes are cyanobacterial photosynthetic energy transfer complexes partly composed of phycobiliproteins, proteins that are widely used as conjugable fluorochromes for flow cytometry. The brightness and photostability of phycobiliproteins suggest that intact phycobilisomes could constitute even brighter probes for fluorescence-based detection systems. Stabilized phycobilisomes have been isolated and the red-excited, far red-emitting Spirulina platensis-derived complex PBXL-3 was accessed as a fluorochrome for flow cytometric immunodetection of surface antigens on immune cells. Although the large size of intact phycobilisomes initially precluded efficient cell surface labeling, the addition of a PEG spacer arm between PBXL-3 and its conjugated avidin molecule (designated PBXL-3L) reduced the steric hindrance associated with the high molecular weight PBXL complex. PBXL-3L increased the surface labeling surface-to-noise ratio and subsequent sensitivity by several-fold over commonly used red-excited fluorochromes such as APC. Interestingly, low power laser sources (including helium-neon and red diode) were particularly efficient at exciting PBXL-3. PBXL-3 was also compatible in with other fluorochromes for multicolor flow cytometry applications. In summary, PBXL-3 was found to possess superior sensitivity and efficiency for flow cytometric immunodetection, particularly with low power laser sources.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Telford WG,Moss MW,Morseman JP,Allnutt FCdoi
10.1016/s0022-1759(01)00367-2keywords:
subject
Has Abstractpub_date
2001-08-01 00:00:00pages
13-30issue
1-2eissn
0022-1759issn
1872-7905pii
S0022175901003672journal_volume
254pub_type
杂志文章abstract：:Radioimmunoassays are usually developed empirically since there have been few established rules which cover general RIA behaviour. Evidence is presented which enables the delineation of 2 types of RIA with distinct group characteristics. (1) Simple RIA, conforming to the law of mass-action kinetics, is a result of uni...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90441-7
更新日期：1985-11-28 00:00:00
abstract：:The sensitivity and specificity of immunohistological staining procedures employing horseadish peroxidase labeled antibody were evaluated with the aid of chessboard titrations. Two indirect staining systems, detecting antinuclear antibody and pemphigus intercellular antibody, were examined with reference to indirect i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90081-8
更新日期：1976-01-01 00:00:00
abstract：:Use of fluorescently labelled multimers, particularly tetramers of peptide and MHC class I glycoprotein (pMHC-I) complexes, is essential for the analysis of CD8+ T cell immunity in basic research and clinical settings. A recently described combinatorial approach using pMHC-I multimers coupled to a unique combination o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.06.003
更新日期：2010-08-31 00:00:00
abstract：:A technically simple microassay for macrophage migration inhibition studies is described. Using simple 0.08 ml of test solution per assay, meaningful statistical analysis via multiple replicates is possible. A novel feature is that test solutions may be stored frozen in the test well until the indicating macrophages a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90252-5
更新日期：1980-01-01 00:00:00
abstract：:Human monoclonal antibodies may be generated by electrofusion of human B lymphocytes with a human/mouse heteromyeloma line. In addition to a fusion protocol optimised for the fusion partners, the activation of B lymphocytes is crucial for fusion and hybrid efficiency. In this study, we initially treated peripheral blo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00431-8
更新日期：2001-09-01 00:00:00
abstract：:The present article reports a multiparametric cytofluorimetric analysis of apoptosis in murine thymocytes and human PBMC from healthy donors or HIV-infected patients. We have evaluated four previously described cytofluorimetric methods of apoptosis quantification, each of them detecting distinct cellular alterations o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00148-2
更新日期：1996-10-30 00:00:00
abstract：:Serologically and biochemically pure HLA antigens and other beta 2-microglobulin-associated proteins were isolated using a rapid immunoprecipitation technique with a monoclonal anti-beta 2-microglobulin reagent A88. Elution of the immunoprecipitate with beta 2-microglobulin isolated from kidney patients proved to be e...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90237-4
更新日期：1981-01-01 00:00:00
abstract：:This report describes a modification of a procedure developed by others for crosslinking IgG to protein A which itself is covalently linked to a gel support. Earlier immunoaffinity columns were described as having large antigen-binding capacities and stability under a variety of elution conditions. The present data sh...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90071-3
更新日期：1990-03-09 00:00:00
abstract：:A study was undertaken to determine whether large amounts of blood (antiserum) could be rapidly removed from rabbits over a two-month period using a simple suction technique. It was found that half a blood volume could be readily removed each week (over 900 ml/rabbit/54 days) without a single fatality. The rabbits eff...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90081-9
更新日期：1978-01-01 00:00:00
abstract：:Human recombinant soluble 37 kDa CD23 has been expressed in insect cells and secreted into the culture medium using the IL-2 leader sequence. The 37 kDa CD23 was purified 600-fold to homogeneity by monoclonal antibody affinity chromatography and gel filtration. The pure protein is monomeric, glycosylated, depleted of ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90253-p
更新日期：1992-05-18 00:00:00
abstract：:By pre-stimulating the lymphocytes of cancer patients with BCGF plus anti-mu chain specific antibody, we have generated human X human hybridomas. The resulting hybridomas preferentially produced IgG class antibody compared with those obtained from non-stimulated lymphocytes. With non-stimulated lymph-node lymphocytes,...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90280-9
更新日期：1990-12-31 00:00:00
abstract：:Complement activation is a key component of the pathogenesis of immune-mediated tissue damage in many diseases. Assessment of complement activation in current practice is largely based on the measurement of intact C3 and C4 or the determination of complement haemolytic function. These parameters reflect activation onl...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(93)90063-d
更新日期：1993-01-04 00:00:00
abstract：:The relative efficiency of depot (Freund's complete adjuvant (FCA) and saponin), and non-depot (Corynebacterium parvum strain P. acnes I, C. parvum strain IMI585, and levamisole) immunoadjuvants were compared in raising agglutinating and precipitating antibodies against the metacestode fluid of Taenia multiceps of she...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90230-2
更新日期：1983-07-29 00:00:00
abstract：:Rabbit antiserum, specific for protein A from Staphylococcus aureus, was conjugated to alkaline phosphatase and used in a double antibody solid-phase enzyme immunoassay. The assay was developed to monitor eluate from a large-scale protein A-Sepharose affinity column used to purify monoclonal antibodies for human clini...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90070-5
更新日期：1985-10-24 00:00:00
abstract：:In transplantation techniques previously described it was impossible to separate the effects of immunosuppressive treatments on the host from the effects directed at the allograft itself. The skin allograft retransplantation technique described here allows one to assess changes in the immunogenicity of the allograft g...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90005-x
更新日期：1984-09-04 00:00:00
abstract：:Ia molecule (Ek,b beta:Ek alpha or Ak beta:Ak alpha)-containing planar membranes were constructed with cholesterol and a 9:1 molar ratio of the phospholipids dipalmitoyl phosphatidylcholine and dilinoleoyl phosphatidylcholine. This lipid composition was found to be optimal for the stimulation of T cell hybridomas of d...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90432-7
更新日期：1987-04-02 00:00:00
abstract：:Mold contamination and exposure to fungi in indoor environments has been associated with various adverse health effects but little is known about the significance of individual fungal species in the initiation or exacerbation of such effects. Using Stachybotrys chartarum as a model fungus we sought to demonstrate that...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2005.12.001
更新日期：2006-02-20 00:00:00
abstract：:Granulocytes and monocytes/macrophages represent key effector cells of the innate immune system. While human monocytes have been recognized as capable of secreting a broad spectrum of cytokines, the situation has been less clear in granulocytes with studies often showing conflicting results. In this study, lipopolysac...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2009.04.001
更新日期：2009-07-31 00:00:00
abstract：:The sandwich solid phase radioimmunoassay procedure has been adapted to the serological diagnosis of tuberculosis by the use of disposable plastic haemagglutination trays in which the wells were sensitized with soluble mycobacterial antigen and the subsequent uptake of globulin from test sera detected with radioiodina...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(75)90069-1
更新日期：1975-01-01 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：: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：:The proinflammatory cytokine IL-18 mediates IFN-gamma production as well as the induction of Th1 polarized immune responses in synergy with IL-12. In this study, we describe the production of isogeneic monoclonal antibodies (Mabs) directed against murine IL-18 (mIL-18). Immunization of IL-18-deficient mice with recomb...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00505-1
更新日期：2002-01-01 00:00:00
abstract：:This work presents a method for analyzing protein microarrays using a colorimetric nanogold probe coupled with silver enhancement (gold-silver detection). In this method, the gold nanoparticles were introduced to the microarray by the specific binding of the gold-conjugated antibodies or streptavidins and then coupled...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.11.008
更新日期：2004-02-15 00:00:00
abstract：:A competitive enzyme immunoassay for rat prolactin using an immunogen and a tracer obtained by recombinant DNA technology is described. Polyclonal antibodies were raised in rabbits immunized with a purified chimeric protein consisting of rat prolactin fused with two synthetic immunoglobulin G binding domains derived f...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90264-x
更新日期：1994-03-10 00:00:00
abstract：:We describe a method for the generation of high-affinity monoclonal antibodies, which combines the power of natural immune responses with in vitro panning, B cell culture, RT-PCR and expression of the recombinant product. B cells from immunised rabbits were incubated at approximately 1000-10,000 cells per well with so...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.08.010
更新日期：2006-10-20 00:00:00
abstract：:Previously we have described studies on in vitro pentamer assembly of Escherichia coli (E. coli) derived heat-labile enterotoxin B subunit (EtxB) using conventional monoclonal antibodies (Amin et al., JBC 1995: 270, 20143-50 and Chung et al., JBC 2006: 281, 39465-70). To extend these studies further we have used phage...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.08.005
更新日期：2008-12-31 00:00:00
abstract：:A sensitive method for staining proteins after transfer from polyacrylamide gels to nitrocellulose paper is described. Transferred proteins are first derivatized by reaction of the nitrocellulose replica with sulfosuccinimidobiotin and are then reacted sequentially with streptavidin, rabbit anti-streptavidin, and hors...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90504-1
更新日期：1986-08-21 00:00:00
abstract：:DNA microarray technology has developed rapidly in recent years and has become an essential tool, providing novel approaches to biomedical research. In this paper, we describe a self-designed ImmunoChip cDNA array for immunological research. With a comprehensive selection of genes of interest, we can focus on key sign...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2005.06.004
更新日期：2005-08-01 00:00:00
abstract：:The goals for the development of multiple antigen peptides (MAP) are to provide a rational and unambiguous system to multimerize different types of synthetic peptide antigens and to attach immunomodulating molecules for targeting and delivery. These goals have been largely realized and new designs of MAPs now permit a...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(96)00066-x
更新日期：1996-09-13 00:00:00
abstract：:The binding of cells to paramagnetic polystyrene microbeads in the absence of coupling antibodies was measured. Cells from normal bone marrow, from an acute lymphoblastic leukemia (ALL) cell line or from a neuroblastoma cell line, were labeled with the fluorochrome Hoechst 33342 and incubated with microbeads by rotati...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90173-7
更新日期：1989-07-26 00:00:00
