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 at exogenous ammonia concentrations of 1000 microM. In the human promyelocytic cell line however, viable cell counts were shown to be reduced at exogenous ammonia concentrations of 300 microM. Next, we determined ammonia and glutamine levels in 11 commercially available media on the day of delivery. It was found that all media contained significantly less glutamine than prescribed. Ammonia was found in all media with concentrations ranging up to 1000 microM. Storage at both 4 degrees C and 20 degrees C caused a further degradation of glutamine and significant accumulation of ammonia in all media. The degradation curves of the various media were used to calculate the first order degradation constant k, which can be used to determine the kinetics of the spontaneous decomposition in culture media. These results suggest that precautions must be taken to avoid the deterioration of commercially available culture media, because of the decay of glutamine. Long storage times lead to a rapid decay of glutamine and an accumulation of the toxic degradation product ammonia.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Heeneman S,Deutz NE,Buurman WAdoi
10.1016/0022-1759(93)90331-zsubject
Has Abstractpub_date
1993-11-05 00:00:00pages
85-91issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(93)90331-Zjournal_volume
166pub_type
杂志文章abstract：:A new sandwich-type solid-phase enzyme immunoassay (EIA) for carcinoembryonic antigen (CEA) was established by using two selected monoclonal antibodies (MAbs). One MAb, F82-61, which is immobilized on polystyrene beads, reacts with an epitope present on the N-terminal domain (N) of the CEA molecule and the other, F11-...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90388-n
更新日期：1993-06-18 00:00:00
abstract：:The Rho family GTPase Cdc42 is a critical regulator of cellular polarization from yeast to man. An analysis of its function in T cell activation is therefore of interest. This analysis poses two substantial challenges, similar to the analysis of many other critical T cell signaling intermediates. First, Cdc42 is requi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.10.017
更新日期：2007-01-30 00:00:00
abstract：:One of the major endpoints of cellular immunological tests remains the quantitative assessment of lymphocyte proliferation as estimated from incorporation of tritiated thymidine into dividing cells. These data are usually generated by beta scintillation counting as strings of counts per minute which require considerab...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90163-a
更新日期：1991-04-25 00:00:00
abstract：:The U937 cell line has been shown to generate tumour necrosis factor alpha (TNF-alpha) in response to soluble stimuli such as PMA and LPS, but only after treatment with GM-CSF. We report here the generation of TNF-alpha from U937 cells following phagocytosis of opsonised zymozan particles without the need for pre-trea...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90141-f
更新日期：1992-08-10 00:00:00
abstract：:In standard displacement flow immunoassays the analyte in the sample creates an active dissociation of labelled antigens (or antigen homologues) from an antigen binding site of an immobilized antibody, after which the labelled substance is measured downstream. Such systems have been described for molecules up to 1 kDa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00106-9
更新日期：1998-08-01 00:00:00
abstract：:A novel enzyme-linked bio-immunoassay (bio-ELISA) has been developed to detect interferon-gamma (IFN-gamma) induced HLA-DR antigen on the surface of human tumor cells. Cells are cultured at 37 degrees C in 96-well microtiter plates in the presence of IFN-gamma for 2 days. After fixation with reagent alcohol the HLA-DR...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90083-5
更新日期：1989-12-20 00:00:00
abstract：:A new radioimmunoassay has been developed for measuring IgM and IgG rheumatoid factors. Diluted sera from donors and patients were incubated with immunoprecipitates prepared from sheep serum and rabbit anti-sheep IgG antiserum. The precipitates were washed, and radioiodinated rabbit F(ab')2 fragments specific from hum...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90259-3
更新日期：1981-01-01 00:00:00
abstract：:Since the number of polymorphonuclear leukocytes (PMN) responding to a chemotactic stimulus in the Boyden chamber is influenced by the cell density, we studied whether this variable was important in determining chemotaxis under agarose. The chemotactic index was determined by summing the product of each cell that had ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90292-1
更新日期：1981-01-01 00:00:00
abstract：:We developed a simple and sensitive enzyme-linked immunosorbent assay (ELISA) for human pulmonary surfactant protein D (SP-D). Human SP-D was purified from bronchoalveolar lavage fluids of patients with pulmonary alveolar proteinosis. Nine monoclonal antibodies (MAbs) were established from BALB/c mice immunized with t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90295-x
更新日期：1994-08-01 00:00:00
abstract：:An ELISA method for the detection of monoclonal antibodies (MAb) to the T3-T cell antigen receptor (TCR) complex was devised. The T3-TCR complex was solubilised using digitonin and a rat anti-T3 MAb (Campath 3) was used to bind it to an ELISA plate. Normal rat serum was used to block cross-reactivity between the rat M...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90280-3
更新日期：1988-01-21 00:00:00
abstract：:A successful technique for the isolation of highly pure suspensions of viable leukocytes from the small intestine of cattle is described. Procedures ranging from mechanical mincing to enzymatic digestion of tissues were compared. The most reliable and reproducible procedure was the sequential treatment of tissues with...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90410-8
更新日期：1987-12-04 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：:Based on the observation that binding of IgM cytophilic antibodies to lymphocytes is temperature dependent, a direct plaque forming cell (PFC) assay was developed to detect IgM-receptor bearing human peripheral blood T and B lymphocytes. Lymphocytes were passively sensitized with IgM anti-SRBC molecules at 4 degrees C...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90330-2
更新日期：1979-01-01 00:00:00
abstract：:An international collaborative study was conducted at ten sites to examine the performance of enzyme immunoassays (EIAs) for the quantitation of IgG1, IgG2, IgG3, IgG4 and total IgG anti-Haemophilus influenzae type b (Hib) capsular polysaccharide in human serum. All groups used the same reagents: microtiter plates coa...
journal_title：Journal of immunological methods
pub_type： 临床试验,杂志文章,多中心研究
doi：10.1016/0022-1759(92)90163-n
更新日期：1992-04-08 00:00:00
abstract：:An in situ immunoassay technique is described which is sensitive to specific binding to surface immobile antigen or antibody. The phenomena of surface plasmon oscillations are shown to be sensitive to antigen-antibody binding on a substrate. An advantage of the technique is that no molecular labeling is required. The ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90295-0
更新日期：1989-06-02 00:00:00
abstract：:Drug-specific monoclonal antibodies (MAbs) were produced against the very small drug hapten (162.15 Da), 5-benzimidazolecarboxylic acid, an analogue of 2-(4-Thiazolyl)benzimidazole (TBZ) but lacking the thiol group. TBZ is widely used as a broad-spectrum anthelmintic in various animal species and humans and also as a ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00340-x
更新日期：2002-12-20 00:00:00
abstract：:Leukemia inhibitory factor (LIF), oncostatin-M (OSM), ciliary neurotrophic factor (CNTF) and cardiotrophin-1 (CT1) act through transmembrane receptors which share the gp190 glycoprotein chain. The understanding of its involvement in the biology of these cytokines is of importance since these systems have recently been...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00074-4
更新日期：1997-07-14 00:00:00
abstract：:Clostridioides difficile (C. difficile) is the most common cause of nosocomial antibiotic associated diarrhoea. The incidence of C. difficile infection (CDI) has been rising worldwide over the last 20 years with consequent rises in morbidity, mortality and healthcare costs, although the incidence has fallen in the UK ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2020.112917
更新日期：2021-01-01 00:00:00
abstract：:Bovine serum albumin (BSA) was used as a non-amphipathic model protein to be included into iscoms. Pretreatment at an acidic pH (2.5) was used to reveal hydrophobic regions, after which BSA could be integrated. In immunization experiments in mice the BSA iscoms induced long lasting and considerably higher serum antibo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90208-d
更新日期：1990-04-17 00:00:00
abstract：:We have developed a modified microtiter plate which has following advantageous features and functions to both conventional microtiter plate and protein array, such as 1) use of conventional microtiter plate reader and washer, and 2) allowance of simultaneous reaction in the same liquid for all wells. Four proteins of ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2007.04.007
更新日期：2007-06-30 00:00:00
abstract：:NIP-conjugated Fab' fragments from a rabbit hyperimmunized with sheep red blood cells (SRBC) were used to link the hapten NIP to target cells for antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent complement-mediated lysis (ADCML). Target cells (SRBC) labelled with this NIP-Fab' complex were c...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90005-z
更新日期：1991-02-15 00:00:00
abstract：:This paper describes a method for processing fresh tissue that allows immunohistological analysis on paraffin sections. The method is based on the use of periodate-lysine-paraformaldehyde fixation. The effects of variation in fixation time, concentration of paraformaldehyde, dehydration, clearing, wax embedding and en...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90106-6
更新日期：1984-12-31 00:00:00
abstract：:Use of whole blood for lymphocyte proliferation experiments have advantages over the usual isolated lymphocyte procedure. We have systematically optimized methodology for whole blood lymphocyte proliferation in male Sprague-Dawley rats. Both phytohemagglutinin (PHA) and concanavalin A (ConA) stimulate lymphocyte blast...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00084-n
更新日期：1995-08-18 00:00:00
abstract：:The advantages of chicken (Gallus gallus domesticus) antibodies as immunodiagnostic and immunotherapeutic biomolecules has only been recently recognized. Even so, chicken antibodies remain less-well characterized than their mammalian counterparts. This review aims at providing a current overview of the structure, func...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/j.jim.2017.05.001
更新日期：2017-08-01 00:00:00
abstract：:A method is described for the purification of monoclonal antibody from mouse ascitic fluid. The fluid is clarified and the lipid removed using silicon dioxide powder, before the immunoglobulin is precipitated using polyethylene glycol. The method provides IgM antibody in high yield and good purity. In the case of IgG ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90483-7
更新日期：1986-07-24 00:00:00
abstract：:Neuraminidase-induced removal of sialic acid from natural substrates (desialylation) unmasks saccharides that are specifically recognized by the lectin peanut agglutinin (PNA). We demonstrate that, when a neuraminidase substrate is coated on to the wells of a microplate, it is possible to quantitate the binding of PNA...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90255-t
更新日期：1990-12-31 00:00:00
abstract：:A new method of mouse B cell isolation has been developed. It is based on the differences in buoyant density of free lymphocytes and cells bound within liposome-cellular complexes (LCC). LCC arise during the interaction of B cells bearing surface immunoglobulins (Ig) with liposomes loaded with affinity purified rabbit...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90074-4
更新日期：1989-12-20 00:00:00
abstract：:A sandwich amperometric enzyme immunoassay with flow injection for alpha-fetoprotein in human serum has been developed with alkaline phosphatase as the enzyme label. p-Hydroxyphenyl phosphate was used as the substrate for alkaline phosphatase. The hydrolysis product, hydroquinone, was detected by oxidative amperometry...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00044-0
更新日期：1996-06-14 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：:We describe a rapid in vitro assay for the evaluation of in vivo properties of conjugate vaccines. Using human and murine monoclonal antibodies (mAb) specific for lipopolysaccharides (LPS), isolated from Pseudomonas aeruginosa, we determined in a competitive binding assay the amount of LPS or conjugate vaccine which w...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90367-5
更新日期：1990-10-19 00:00:00
