Abstract：
:We have developed a solid-phase assay for the quantification of terminal deoxynucleotidyl transferase (TdT) enzymatic activity in crude cellular extracts. Affinity-purified, polyclonal anti-TdT antibodies are bound to the wells of a microtiter plate, and TdT in extracts is then bound to the immobilized antibodies. The enzymatic activity of the antibody-bound TdT is measured directly in the wells of the microtiter plate. This method yields highly reproducible values, even with samples of low activity. Because it is also technically very simple, it is ideal for determining enzymatic activity for large numbers of clones with limited numbers of cells.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Harriman W,Fischer ER,Wabl Mdoi
10.1016/0022-1759(95)00004-tsubject
Has Abstractpub_date
1995-04-26 00:00:00pages
221-4issue
2eissn
0022-1759issn
1872-7905pii
002217599500004Tjournal_volume
181pub_type
杂志文章abstract：:A proliferation assay to detect a specific T cell response against HLA class I-binding peptides is reported. To establish the specificity and sensitivity of the assay we used a synthetic peptide derived from the CMV glycoprotein B containing the HLA-A2.1 ligand motif. Lymphocyte proliferation was measured following cu...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00124-x
更新日期：1996-10-16 00:00:00
abstract：:Surface plasmon resonance is a valuable optical phenomenon for monitoring biomolecular interactions in real time. In this project anti-mouse-Fc was coupled to the carboxymethyl dextran coating on the surface of a CM5 sensor chip (BIAcore) using amine coupling. Monoclonal antibodies (MAbs) to Luteinizing Hormone (LH) w...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00167-7
更新日期：1998-12-01 00:00:00
abstract：:High-titered antisera against human melanoma- and leukemia-associated antigens have been produced in non-human primates. The data from this and from other studies suggest that this model may prove a valuable source of highly specific antisera against a variety of human tumor-associated antigens or alloantigens. Our me...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90048-x
更新日期：1976-01-01 00:00:00
abstract：:Accurate detection of apoptotic cells is important for the determination of cell viability. The aim of this study was to compare the sensitivity of the cell permeant SYTO 16 fluorescent dye for detecting early apoptotic mononuclear cells (MNCs) in normal donor blood with other apoptosis assays [i.e. Annexin-V, light s...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2005.07.017
更新日期：2005-10-30 00:00:00
abstract：:The diagnostic and therapeutic applications of antibody single-chain Fv (sFv) fragments often require large amounts of protein that can be problematic and expensive to obtain. Here we report the secretion of two sFv fragments by the yeast Pichia pastoris at levels up to 250 mg/l. Soluble sFv fragments were purified fr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00213-x
更新日期：1997-02-14 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
abstract：:Five beta-diketone derivatives were studied for multiple labelling of proteins. The labelled proteins were characterized by absorption and fluorescence measurements. It was found that proteins labelled with chlorosulfonyl-thenoyltrifluoroacetone (CTTA) were able to form highly fluorescent complexes with Eu3+ which exh...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00289-9
更新日期：1995-02-27 00:00:00
abstract：:A system proposed for measurement of mutational risk consists in detection of hemoglobin mutations expressed in erythrocytes. For this detection the production of antibodies specific for Hb variants is essential. Recently we reported a sensitive solid-phase EIA for the production and selection of polyclonal and monocl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90189-8
更新日期：1983-05-13 00:00:00
abstract：:Lymphocyte subsets from human blood obtained by different procedures were analyzed for cytotoxic potential and phenotypic characteristics. Nylon wool column passed lymphocytes were fractionated on the basis of: (1) E and Fc gamma receptor expression, (2) cell density and Fc gamma receptor expression, (3) Fc gamma rece...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90209-0
更新日期：1983-09-30 00:00:00
abstract：:An antibody detection ELISA was developed for diagnosis of visceral leishmaniasis. Antigens released by Leishmania donovani promastigotes into a protein-free medium were used. SDS-PAGE analysis has indicated that Ld-ESM contain several protein antigens. Titration and chequer-board analyses were performed to optimise t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00341-6
更新日期：2001-06-01 00:00:00
abstract：:Human vascular endocan is a proteoglycan exhibiting tumorigenic activity through both its glycan and protein cores. Endocan mRNA is identified as being one of the most significant molecular signatures defining a poor prognosis in lung, breast, kidney, prostate, and thyroid malignancies. The survival inversely correlat...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.02.009
更新日期：2012-04-30 00:00:00
abstract：:The majority of vaccines and several treatments are administered by intramuscular injection. The aim is to engage and activate immune cells, although they are rare in normal skeletal muscle. The phenotype and function of resident as well as infiltrating immune cells in the muscle after injection are largely unknown. W...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.06.011
更新日期：2015-10-01 00:00:00
abstract：:A paper radioimmunosorbent test (PRIST) was shown to be sensitive and reproducible when used with excretory/secretory antigen of Toxocara canis second stage larvae. Whatman No. 50 filter paper (5 mm discs) gave the most consistent and clear results with antigen at a concentration of 100 micrograms/ml, and could be sto...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90180-5
更新日期：1980-01-01 00:00:00
abstract：:Phage-displayed random peptide libraries are important tools in identifying novel epitopes/mimotopes that may lead to the determination of antigen specificity. In this approach, high-affinity phage peptides are enriched by affinity selection (panning) on a monoclonal antibody. To facilitate identification of all poten...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.08.006
更新日期：2006-10-20 00:00:00
abstract：:A competitive enzyme immunoassay with labelled antibodies has been developed for methotrexate (MTX). Methotrexate in the sample and a constant quantity of this hapten physically absorbed to polystyrene spheres through a methylated bovine albumin carrier were allowed to compete for a limiting amount of peroxidase label...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90353-8
更新日期：1983-05-27 00:00:00
abstract：:We developed a reliable method for reconstituting thymectomized, lethally irradiated guinea pigs. Injection of 2.5-10 X 10(7) syngeneic bone marrow cells into adult thymectomized, lethally irradiated guinea pigs produced survival of 46-100% of treated animals. Gentamycin sulfate (5 mg/kg of body weight) for 10 days wa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90187-x
更新日期：1984-05-25 00:00:00
abstract：:A method of obtaining rat thoracic duct lymphocytes is described, including a review of the surgical technique, with several modifications, and including an outline of a newly designed rat restrainer that helps to triple thoracic duct output compared to the Bollman restrainer. The new restrainer allows the rat increas...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90101-9
更新日期：1988-06-13 00:00:00
abstract：:Melanins are notoriously difficult to work with because of their unique physical and chemical properties. The study of melanins is hampered by the scarcity of melanin-specific reagents and serological techniques. In this study we describe modifications to the standard method for the isolation of melanins from in vitro...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00255-6
更新日期：2000-10-20 00:00:00
abstract：OBJECTIVE:The central mechanism for establishing a self-tolerant and functional T cell repertoire includes the promiscuous expression of otherwise tissue-restricted proteins by medullary thymic epithelial cells (TEC). We here demonstrate a novel and highly efficient method for isolating this rare key cell type. METHOD...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2019.02.001
更新日期：2019-04-01 00:00:00
abstract：:The chemiluminescence (CL) assay has been used to measure the reactive oxygen species (ROS)-generating capacity of phagocytes. To achieve more optimal measurement conditions for a multi-channel microplate photon-counting CL analyzer with the cooled charge-coupled device (CCD) camera which offers enhanced sensitivity, ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00159-2
更新日期：1997-12-15 00:00:00
abstract：:An alternate method was designed to couple a limited quantity of protein to an affinity support when a conventional technique was unsuccessful. This was achieved through the introduction of a small number of sulfhydryl groups to the ligand by reaction with 2-iminothiolane which resulted in a limited number of reactive...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90034-8
更新日期：1988-08-09 00:00:00
abstract：:We report the development of an expression system for the production of soluble, calmodulin (CaM)-tagged proteins in Drosophila Schneider S2 cells and the subsequent use of these proteins for the selection of phage displayed antibodies. The CaM-tag permitted the purification of recombinant protein to >90% purity in a ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.08.003
更新日期：2006-10-20 00:00:00
abstract：:The affinity of guinea pig thymocytes and peripheral T lymphocytes for rabbit erythrocytes was found to be enhanced following treatment of the erythrocytes with papain. By increasing the numbers of rosette-forming cells and giving stronger, more stable rosettes, this procedure increases the usefulness of the reaction ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(75)90013-7
更新日期：1975-06-01 00:00:00
abstract：:Many enzyme-linked immunosorbent assays take advantage of immobilized antigens for the identification of antibody binding sites. Generally, the analysis of cellulose membrane-bound B-cell epitopes is currently considered of high utility. We adapted this methodology for the stimulation of a T helper cell hybridoma with...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00194-5
更新日期：2000-01-13 00:00:00
abstract：:The fluorescence activated cell sorter (FACS) was used with an indirect membrane immunofluorescence technique to detect antibody against the Forssman antigen, a glycosphingolipid. Sheep erythrocytes, which contain Forssman antigen as a major membrane glycosphingolipid, were used as the target antigen. Detection of the...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90098-4
更新日期：1983-02-25 00:00:00
abstract：:There is an emerging trend in the pharmaceutical industry to evaluate a variety of surrogate biomarkers in Phase I/II clinical studies with the intention of determining potential activity of drugs early in clinical development. A number of cytokines expressed in pathological conditions are currently being considered a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00543-9
更新日期：2002-02-01 00:00:00
abstract：:Certain serum samples produce high background in Western and direct immunoblot assays that detect human serum IgG against specific antigens. We determined that this was due to a reaction between endogenous IgG and the membrane blocking agent (we refer to this as blocking-specific background). Using milk as blocking ag...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90259-a
更新日期：1993-01-14 00:00:00
abstract：:A scheme for quantitation of serum and urinary paraproteins is described using isoelectric focusing and scanning densitometry. Paraproteins could be quantified using this system when present at concentrations ranging from 1 mg/ml to 26 mg/ml, depending on the immunoglobulin class. The relevance of these results to mon...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90082-7
更新日期：1986-06-24 00:00:00
abstract：BACKGROUND:Accumulating evidence intensively advises circulating endothelial progenitor cells (EPCs) and monocyte subsets as surrogate cellular biomarkers in cardiovascular and cancer disease. However, a general standard on their quantification is still elusive, thus precluding a routine monitoring and comparative inte...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.04.003
更新日期：2012-07-31 00:00:00
abstract：:An Ia-rosette assay for the detection of Ia-like alloantigens on the chicken lymphoid cell surface is described. The method is based on the ability of cells treated with alloantiserum to Ia and then with rabbit antiserum against the Fc portion of chicken IgG, to form rosettes with sheep erythrocytes (SRBC) coated with...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90161-7
更新日期：1981-01-01 00:00:00
