Abstract：
:A competitive RT-PCR method that permits reliable quantification of minute amounts of reverse-transcribed mouse lymph node mRNA is described. Using this technique, an absolute number of cDNA copies ranging from 10(3) to 10(5) can be determined, with a precision superior to 25%. The standard templates described in the present study permit the quantitation of beta-actin, IFN gamma, IL2, IL3, IL4, IL10, IL12 (p40 subunit), TGF beta 1, inducible nitric oxide synthase, ELAM-1, VCAM-1, and ICAM-1 mouse mRNA. The expression of a particular transcript is normalized to an arbitrary number of actin transcripts. The standard templates and wild-type cDNA have nearly identical sequences, but they can be distinguished by unique restriction sites. Known amounts of these standard templates, are co-amplified with serial dilutions of the cDNA derived from the mRNA of interest. Oligonucleotide primer pairs possessing 3' octamers found infrequently in the mouse genome (< or = 0.26 x 10(-6)) are used to amplify sequences, chosen to contain no GC stretches longer than 8 (PCRare software) (Griffais et al., 1991). Samples of each PCR product are digested separately with restriction endonucleases unique either for the wild-type or the standard amplicon. The quantitation of the test product and the standard product is easily carried out following their electrophoresis in an ethidium bromide-stained agarose gel.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Colle JH,Falanga PB,Singer M,Hevin B,Milon Gdoi
10.1016/s0022-1759(97)00186-5subject
Has Abstractpub_date
1997-12-29 00:00:00pages
175-84issue
2eissn
0022-1759issn
1872-7905pii
S0022-1759(97)00186-5journal_volume
210pub_type
杂志文章abstract：:Choosing optimum pair of capturing antibody and detecting antibody when developing monoclonal antibody (MAb)-based, sandwich enzyme-linked immunosorbent assays is a time-consuming process requiring the coupling of individual antibodies to an enzyme like horseradish peroxidase or alkaline phosphatase. The MAbs required...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00287-1
更新日期：1996-05-10 00:00:00
abstract：:Signalling pathways modulated by the family of IL-1/TLR receptors are central to innate immune responses. Novel components playing key regulatory roles in these pathways continue to be isolated. Here we describe the use of autocatalytic vectors for identifying critical components of these signalling pathways. The meth...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2007.11.002
更新日期：2008-01-31 00:00:00
abstract：:Western blotting is one of the most commonly and widely used methods for identifying the presence of an antigen in biochemical studies. The present study describes a novel 'coupled blotting' approach that allows antigen on dot and Western blots to be simultaneously probed with primary and secondary probing reagents. T...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00102-0
更新日期：1996-09-09 00:00:00
abstract：:The neutrophilic granulocyte is the most numerous leukocyte in peripheral blood. The development from a multipotent progenitor cell to a mature neutrophil takes place in the bone marrow over a period of 10-14 days. In order to understand the cellular mechanisms behind this process, it is necessary to investigate cells...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(99)00176-3
更新日期：1999-12-17 00:00:00
abstract：:IL-7 is a crucial cytokine for T cell hematopoiesis and peripheral homeostasis which by signaling through its receptor alpha chain (CD127) is essential for inducing T cell proliferation and survival. Since the specific CD127 alpha chain is found in a soluble state (sCD127) and at a high level in plasma (ng/mL), it is ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2009.12.003
更新日期：2010-02-28 00:00:00
abstract：:Irradiated human peripheral blood lymphoid cells undergo apoptosis and progressively exhibit typical changes in light scatter and plasma membrane integrity that can be easily tracked by flow cytometry. Using this model, we assessed the capacity of a newly developed fluorochrome, Apostain, in identifying early apoptosi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00054-9
更新日期：1997-06-23 00:00:00
abstract：:Sepsis is a systemic inflammatory response ensuing from presence and persistence of microorganisms in the bloodstream. The possibility to identify them at low concentrations may improve the problem of human health and therapeutic outcomes. So, sensitive and rapid diagnostic systems are essential to evaluate bacterial ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.12.004
更新日期：2019-02-01 00:00:00
abstract：:Different Percoll density gradients were used to purify granulocytes from Long-Evans, Fischer, Sprague-Dawley, and Fischer/Long-Evans hybrid rats. Three different discontinuous gradient types were developed which permitted the separation of polymorphonuclear leukocytes (PMNLs) from the peripheral blood of the differen...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90093-0
更新日期：1994-01-03 00:00:00
abstract：:Natural killer (NK) cells are important mediators of virus- and tumor-specific immune responses. The transfection of genes into NK cells has been proven difficult and so far requires infection with virus-based vectors. Here, the application of a novel nonviral, electroporation-based gene transfer method is described f...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00431-3
更新日期：2003-03-01 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：:Pertussis toxin is known to elicit lymphocytosis in whooping cough patients and experimental animals, by blocking the extravasation of lymphocytes and stimulating their release from lymphoid organs such as the thymus. The mechanisms responsible for these unique effects of PT are not fully understood. The effect of per...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90090-3
更新日期：1991-11-22 00:00:00
abstract：:In developing countries the acquisition and use of radioisotopes with their short shelf lives poses enormous problems. The objective of this study was to develop a simple, sensitive, non-isotopic method for the measurement of TSH in human serum. A conventional sandwich enzyme-linked immunosorbent assay (ELISA) techniq...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90315-8
更新日期：1988-11-25 00:00:00
abstract：:Enzyme-linked immunoadsorbent assay (EIA) has widespread use for the measurement of antibody concentration. The affinity constant (Kaff) of the antibody has an effect upon the quantification by EIA. It is thus important to be able to measure Kaff by solid-phase EIA. Based upon the Law of Mass Action and using serial d...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90187-6
更新日期：1987-06-26 00:00:00
abstract：:We developed a noncompetitive two-site sandwich ELISA to quantitate monoclonal antibodies in culture supernatant. This assay measures the initial enzyme activity rate during the first minute of the reaction, which ensures linear velocity relative to time and a progress curve slope proportional to analyte concentration...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2019.112645
更新日期：2019-11-01 00:00:00
abstract：:A novel bioanalysis system based on immunochromatography was developed in connection with a nitrocellulose resin modified micropipette tip, such as ZipTip. The sandwich-type immunoassay was applied to our bioanalysis system. The simple handling of the micropipette enabled us to increase the sample volume and detect lo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.02.011
更新日期：2006-05-30 00:00:00
abstract：:The lysis of susceptible targets by efficient cytotoxic T lymphocytes (CTL) increases both with time and with the ratio of CTL to target. Simple methods for calculating a killing rate constant from the time dependence of killing and for calculating the relation of the killing rate constant to the concentration of exoc...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00009-9
更新日期：1996-06-10 00:00:00
abstract：:Bromodeoxyuridine (BrdU) is used to label synthesizing DNA and to chase label-retaining cell (LRC). As stem cells divide slowly in adult tissues, they can be visualized as LRCs. In order to identify LRCs in hard tissue, we examined optimal conditions of fixation, demineralization, and DNA denaturation/antigen retrieva...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.07.013
更新日期：2008-11-30 00:00:00
abstract：:We have developed a microarray (DotScan) that enables rapid immunophenotyping and classification of leukaemias and lymphomas by measuring the capture of cells by immobilized dots of 82 CD antibodies [Belov, L., de la Vega, O., dos Remedios, C.G., Mulligan, S.P., 2001. Immunophenotyping of leukemia using a cluster of d...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2005.07.007
更新日期：2005-10-20 00:00:00
abstract：:Antigenic epitopes recognized by FoxP3(+) regulatory T cells (Treg) are poorly defined, largely due to a lack of assays for determining Treg specificity. We have developed a novel approach for detecting human Treg specific to peptide antigen, utilizing down-regulation of surface CD3 as a read-out of antigen recognitio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.01.004
更新日期：2012-03-30 00:00:00
abstract：:A simple method is described for the preparation of vimentin-type intermediate filaments for solid-phase immunoassays. Bovine aortic endothelial cells were grown to confluency in multiwells, and extracted with Triton X-100 and a high-salt buffer. Residual protein in the wells consisted of 53% vimentin as judged by SDS...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90149-8
更新日期：1985-12-27 00:00:00
abstract：:We describe here a specific and sensitive assay for biologically active bovine type-I interferon (IFN) in an Mx/CAT reporter gene assay. The assay is based on Madin-Darby Bovine Kidney cells transfected with a plasmid, containing a human MxA promoter driving a chloramphenicol acetyltransferase (CAT) cDNA. CAT expressi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00359-8
更新日期：2001-03-01 00:00:00
abstract：:A method for bleeding rats from the tail vein was developed. Sufficient vasodilation of the tail was accomplished by the use of a specially designed warming device. With the help of a physical restrainer, up to 5 ml of blood can be collected from the tail vein and no anaesthesia is required. This bleeding procedure is...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90148-x
更新日期：1982-09-17 00:00:00
abstract：:A new rapid staining and measuring method has been developed for the quantification of migrated cells in a microchemotaxis chamber. The migrated cells were, after staining, evaluated by a transmission densitometer. The method introduced here is more accurate and faster than those described previously. In addition the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90051-3
更新日期：1989-03-10 00:00:00
abstract：:5-Bromo-2-deoxyuridine (BrdUrd) is a thymidine analogue which can be used to detect DNA synthesis in cells. The present paper describes a method for the fixation and DNA denaturation of cells that affects neither the morphology of the cells not their adhesion to a glass surface. The cells were fixed in absolute ethano...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90401-2
更新日期：1988-04-06 00:00:00
abstract：:Factors influencing the performance of a dipstick competitive particle capture immunoassay (PCI) for the steroid oestrone sulphate (OS) were investigated. Appropriate 'blocking' of the nitrocellulose dipstick membrane was necessary for the upward flow of microsphere particles. Traditional protein blocking agents inclu...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00280-6
更新日期：2002-12-01 00:00:00
abstract：:Wegener's granulomatosis (WG) is a systemic vasculitis which is diagnosed on clinicopathological findings. The diagnosis may be aided by the presence of anti-neutrophil cytoplasm antibodies (ANCA). In WG, ANCA are primarily directed to proteinase 3 (PR3), a serine protease of the azurophilic granules of the neutrophil...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90364-6
更新日期：1994-10-14 00:00:00
abstract：:A microtitre plate enzyme immunoassay (EIA) for determining low concentrations of IgA is described and validated for serum and plasma products. The measuring range of the assay was 3.3-150 micrograms/l. The predilution requirement of samples was matrix dependent, ranging from 1/16 for serum to 1/2 for 4% albumin. Dilu...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90239-4
更新日期：1993-07-06 00:00:00
abstract：:Electrochemiluminescence (ECL) assays have been widely used for the detection of anti-therapeutic antibodies (ATAs) against biotherapeutics. With the discontinuation of BioVeris (BV) ECL platform, an alternative technology was needed to replace BV assays to ensure continuous support of multi-year clinical studies. Aft...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.09.013
更新日期：2010-10-31 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：:We describe optimal conditions for protein labeling with a new fluorescent probe, 4,7-bis-(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA). The labeled proteins are suitable for time-resolved fluorometric applications because BCPDA forms fluorescent complexes with Eu3+ which exhibit very long fluo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90031-2
更新日期：1988-08-09 00:00:00
