Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Sensitive identification of human blood and the determination of ABO blood group from a minute bloodstain were simultaneously carried out by a direct ELISA-ABC method. A cotton thread (1 cm in length) stained with 1 microliter of human or animal blood was stored for 2-4 weeks at room temperature. Hemoglobin (Hb) of the bloodstained thread was gently extracted with 100 microliters of PBS at room temperature, and the thread was washed with PBS to dehemoglobinize. And ABH blood group antigens were extracted from the same dehemoglobinized thread with 100 microliters of 5% ammonia solution at 56 degrees C. The extracts of PBS and ammonia were two-fold serially diluted with 0.1 M sodium carbonate buffer, coated to the wells of a flat bottomed microplate. The PBS extract was tested with a biotinylated antibody against human HbA0 for identification of human blood. Human blood was clearly distinguishable from bloods of other species including Japanese monkey. The minimum detection limit of human blood of the PBS extract of the bloodstained thread was 1:40,960 (3.4 ng Hb), and the limit was found to be approximately 200 times higher than that obtained by a leucomalachite green test or by a precipitation ring test using anti-human HbA serum. The ammonia extract was tested with biotinylated anti-A, anti-B and anti-H antibodies for ABO blood grouping. ABH antigens of the ammonia extract of the bloodstained thread were clearly detected. The minimum determination limits of blood group A, B, AB and O of the ammonia extracts were 1:160, 1:160, 1:80 and 1:160, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Sensitive identification of human blood and simultaneous determination of ABO blood group from a minute bloodstain by an ELISA-ABC method]. 130 54

The binding of MoAbs and PoAbs to native GnRH, azo-GnRH, GnRH-BSA and azo-GnRH-BSA conjugates was studied using a solid phase ELISA test. The use of carbonate/bicarbonate buffer (pH 9.2) significantly affected the coating of peptide to the ELISA plates. Azo-GnRH coated in PBS showed seven times less binding to MoAb than the native GnRH. Saturation analysis studies indicated comparable binding to the antibodies to the coated GnRH and GnRH-BSA but binding to azo-GnRH-BSA was higher than to azo-GnRH. The epitope recognition ability of MoAbs was therefore investigated. Blocking ELISA additivity tests were performed to analyse whether MoAbs recognised a common epitope involving a conformation or sequence of the modified peptide (azo-GnRH). The plates coated with azo-GnRH or azo-GnRH-BSA and saturated with MoAbs did not bind PoAbs. The inhibition activity was similar when GnRH or GnRH-BSA was used instead of azo-GnRH or azo-GnRH-BSA thus suggesting that the MoAbs were directed against a conformational epitope of the GnRH molecule.
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PMID:Influence of immunogen modification of the gonadotropin releasing hormone antibody reactivity. 138 94

Conditions of the enzyme immunoassay for the detection of HSV-I by the "sandwich"-method at all stages of the testing were optimized using monoclonal antibodies (monoAB). When identical concentrations of monoclonal antibodies and durations of their adsorption on polystyrene plates were used, the signal in EIA (A405) with carbonate buffer was by 30% higher than that for PBS. A similar increase in the signal was observed at a temperature of incubation of 37 degrees C as compared with that at 4 degrees C. Even during 18-hour incubation at 37 degrees C no inactivation of monoAB occurred. It was further shown that saturation of the surface of wells with protein was achieved at a concentration of monoAB 5-10 micrograms/ml, and under these conditions the optimal dilution of the conjugate was that corresponding to antibody concentration of 1.5 micrograms/ml. When ABDC peroxidase and 5-aminosalycilic acid were used as the substrate, the presence of HSV-I was regularly detected even with a concentrated virus preparation diluted 500-1000-fold (corresponding to approximately 5.0 1g TCID50).
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PMID:[Development of an immunoenzyme test for determining herpes simplex type 1 virus using monoclonal antibodies]. 303 3

The effect of prostaglandin E2 (PGE2), on the intracellular pH (pHi) in BCECF-loaded Madin Darby Canine Kidney (MDCK) cells was investigated. PGE2 elevated the pHi. Under resting conditions, pHi of MDCK cells suspended in PBS at pH 7.4 was 7.11 +/- 0.08; PGE2 increased pHi with an EC50 of 0.16 microM. PGF2 alpha elicited a similar response to PGE2, with an EC50 of 0.24 microM. Amiloride (0.4 mM) reversed the response to PGE2 (control 7.18 +/- 0.05; PGE2 7.26 +/- 0.05; after amiloride 7.18 +/- 0.05). In MDCK cells exposed to a Na(+)-free solution, alkalinization induced by this eicosanoid was blocked (Ringer-choline 7.16 +/- 0.03; PGE2 7.16 +/- 0.02). PGE2 increased by 100% the rate of recovery after an acidification pulse with ammonium chloride. In the presence of Ringer-HCO3- (pH 7.4), there was a delay in the maximal response to this prostaglandin (PBS 2.2 +/- 0.27, Ringer-bicarbonate 3.4 +/- 0.55 min) and the pHi increment was less marked than in PBS (0.09 pH units in HCO3- versus 0.16 pH units in PBS; P < 0.001). This effect of PGE2 was not blocked by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (1.0 mM). PMA (100 nM), activator of protein kinase C, mimicked the response to PGE2, suggesting the participation of this kinase on the effect of the prostanoid. As expected, two inhibitors of protein kinase C, staurosporine and sphingosine, abolished the response to PGE2. Staurosporine (0.10 microM), an inhibitor of protein kinase C, blocked the response to PGE2 (control 7.02 +/- 0.04; PGE2 and staurosporine 7.03 +/- 0.04, n = 9, not significant). Sphingosine, another inhibitor of protein kinase C, also blocked the response to PGE2. Two analogues of cAMP did not modify the pHi. In summary, PGE2 induced an intracellular alkalinization via stimulation of a Na+/H+ exchanger, with the participation of protein kinase C, in MDCK cells.
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PMID:Induction of alkalinization in cultured renal cells (MDCK line) by prostaglandin E2. 748 Jul 99

Water-soluble extracts from psoriatic scales and normal human skin were prepared using either phosphate-buffered saline, pH 7.2, or 0.1 M carbonate buffer, pH 10.8. Anaphylatoxin C5a des Arg was quantified using a novel sandwich enzyme-linked immunosorbent assay (ELISA) using neoepitope-specific monoclonal antibodies. Alkali was about five to eight times more efficient than PBS in extracting C5a des Arg from scales, probably via dissociation of bound C5a des Arg. C5a des Arg concentration in scales from three patients suffering from psoriasis vulgaris varied between 2.5 and 4.6 ng/mg scale. No C5a des Arg was detectable in normal skin extracts. The biological activity of alkali-extractable C5a des Arg, i.e. chemotaxis, was preserved. The concentration of C5a des Arg relative to the concentration of albumin was taken as a parameter of the degree of complement activation in the psoriatic lesions, and was found to be more than six times higher than values attained in serum after maximum complement activation by zymosan. We conclude that complement activation may play a quantitatively important role in the inflammatory process in psoriasis.
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PMID:Surprisingly high levels of anaphylatoxin C5a des Arg are extractable from psoriatic scales. 850 93

The characterisation and selection of membranes by means of an immunofiltration assay is described. The chemical composition of the membranes was: nitro-cellulose, polyamide, polyvinylidene difluoride, polyethersulfone, cellulose acetate, regenerated cellulose, cellulose nitrate, and glass fibre. In order to characterise the membranes according to their binding capacity, immobilisation stability, sensitivity and hydrodynamic properties, two basic immunofiltration formats were performed. In both formats, enzyme label (horseradish peroxidase, HRP) and colorimetric detection were used. In the immobilised antibody format, three monoclonal antibodies (mAb) against the insecticide carbaryl were immobilised on the membranes by passive adsorption. In the immobilised hapten format, two haptens conjugated to bovine serum albumin (BSA) were immobilised. Immobilon-P was the best membrane with regard to the characterisation criteria and permitted the filtration of large volume (5.0 ml) through the membrane without release of the receptor. The immobilisation of the receptor (antibody or haptenic conjugate) was pH dependent. Good results with regard to mAb-antigen recognition, were obtained using 50 mM carbonate/bicarbonate buffer, pH 9.6. However, the most sensitive assays were achieved using, 10 mM phosphate buffer, 137 mM NaCl, 2.7 mM KCI (PBS), pH 7.4 as immobilisation buffer. Furthermore, all these results permit the choice of the best membrane for the rapid and sensitive determination of carbaryl. This study will assist the development of dipsticks, immunoelectrodes, membrane-based immunoreactors or immunoconcentration devices that are based on the use of membranes as immunosupports.
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PMID:Selection and characterisation of membranes by means of an immunofiltration assay. Application to the rapid and sensitive determination of the insecticide carbaryl. 1035 11

Specific monoclonal antibodies (MAbs) to mefloquine conjugated to bovine serum albumin (mefloquine-BSA) were produced by hybridoma technology. The mefloquine-BSA was synthesized by converting mefloquine into hemisuccinate followed by convalently linked to bovine serum albumin (BSA) and coupling with N,N' disuccinimidyl carbonate (DSC). The conjugate was purified by Sephadex G-75 gel filtration using 0.01 M PBS pH 7.2. An average of 19.34 molecules of mefloquine were conjugated to each molecule of protein determined by differential UV absorption spectra of hapten and protein carrier. Sixteen monoclones producing antibody specific to mefloquine were screened by indirect ELISA using homologous antigens. The specificity of MAbs was determined by reacting with BSA and the structurally related antimalarial drug, quinine. Three, three, five and two MAbs belonged to IgG1, IgG2a, IgG2b and IgG3, respectively. Most of the MAbs slightly reacted with quinine-BSA due to the closely related structure of mefloquine to quinine. The selected MAb designated 11F9(G5)G9 which showed no cross reaction with quinine-BSA gave high reactivity with blood samples from malaria patients previously treated with mefloquine when compared to normal blood by indirect ELISA. The preliminary results indicated that such specific MAb could be used as antibody probe for detection of mefloquine in biological fluids.
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PMID:Characterization of specific monoclonal antibodies for detection of mefloquine in body fluids. 1128 97

Bioresorbable polymers have found a wide range of uses in medical implants, from sutures to scaffolds for tissue engineering applications. Increasingly they are being used in internal orthopaedic fixation devices, in which the strength retention profile is important. Polyglyconate B, a block co-polymer of glycolic acid and trimethylene carbonate, is one polymer used in these applications. In this study, the hydrolytic degradation of polyglyconate B has been studied in vitro. Specimens were prepared with two initial molecular weights and aged in PBS solution at 37 degrees C for up to 31 days. The polymers were characterised by gel permeation chromatography. for molecular weight, tensile testing and mass change, as a function of degradation time. A further aim of the work was to determine whether the measured changes in tensile strength over time could be fitted to a simple model. The results showed that the observed relationship between strength and molecular weight was more complex than that used in our model. However, the data could be modelled using an empirically derived relationship between tensile strength and number average molecular weight (Mn). Changes in other mechanical properties, such as strain at break, were also found to be strongly dependent on changes in the single parameter of Mn.
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PMID:Hydrolytic degradation of polyglyconate B: the relationship between degradation time, strength and molecular weight. 1216 96

Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciola hepatica, peroxidase of conjugate anti-cattle IgG and orthophenylenediamine as a substrate by micro-method technique of Voller et al. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows: In assay for optimal dilution of stock antigen, the antigen (protein contents; 0.8 mg/ml) was diluted from 1/50 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y=-0.181-0.00127X in infected sera, and log Y= -0.578- 0.000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. In assay for optimal dilution of sera, the sera were diluted from 1/25 to 1/400 with in PBS/ Tween 20(pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y=-0.1540-0.0007238X in infected sera and log Y=-0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). In the 27 cases of negative intradermal test, OD values of the ELISA are 0.447 +/- 0.144, the 95 percent confidence interval (Mean + 2 x SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100 percent in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was 0.846 +/- 0.224). The 75 percent (44 cattle) among them had higher value with compared to the criterion, and the 60 percent (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. Prevalence of fascioliasis was 43.4 percent in the application of the ELISA to 272 cattle which were reared in Jeonbug district.
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PMID:[Application of micro-ELISA in serodiagnosis of fascioliasis in cattle] 1288 91

The objective of this study was to investigate the method and effect of blocking the specific reaction between lymphocyte HLA-I antigen and its antibody. The lymphocytes were disposed with 12 mmol/L methoxypolyethelene glycol benzotriazol carbonate (mPEG-BTC) in concentration gradient in PBS (pH 7.4) at 22 degrees C. The effect of the modified lymphocytes was detected by microlymphocytotoxicity assay. The results showed that lymphocytes modified by mPEG-BTC did not react with related HLA-I antibodies in microcytotoxicity test. It is suggested that the specific reaction between HLA-I antigen of lymphocyte and HLA-I antibodies can be completely camouflaged by mPEG-BTC in PBS (pH 7.4) under 22 degrees C room temperature.
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PMID:[Camouflage of HLA-I antigen in lymphocyte surface]. 1470 47


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