Radioimmunoassay for free and conjugated urinary metanephrine.

1977 ◽  
Vol 23 (7) ◽  
pp. 1264-1267 ◽  
Author(s):  
R W Lam ◽  
R Artal ◽  
D A Fisher
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Incubation Time ◽  
Bovine Serum ◽  
Normal Subjects ◽  
Specific Radioimmunoassay ◽  
Coefficients Of Variation ◽  
Assay Tube ◽  
Before And After ◽  
Urine Specimens

Abstract We describe a convenient and specific radioimmunoassay for urinary metanephrine, in which we used an antiserum generated in rabbits by injecting with synephrine conjugated to bovine serum albumin as described by Grota and Brown [Endocrinology 98, 615 (1976)]. The antiserum is specific for metanephrine and epinephrine. Epinephrine cross reaction is minimized to 1% by adding 2 ng of epinephrine to each assay tube. Incubation time is 12 h. The sensitivity of the assay is 40 pg. Within-and between-assay coefficients of variation are 7.0 and 9.7%, respectively. We assayed first-voided morning urine and 24-h urine specimens from 15 normal subjects before and after acid hydrolysis, to assess total and free metanephrine concentrations. Mean (+/-SD) total and free urinary metanephrine excretion were 17 +/- 11 and 3.5 +/- 1.0 micrograms/24 h in these subjects; corresponding values for the morning specimens were 13.4 +/- 8.3 and 3.5 +/- 1.5 micrograms/liter, respectively.

A COST REDUCTIVE METHOD FOR THE RADIOIMMUNOASSAY OF PLASMA ANDROSTENEDIONE. A COMPARISON OF ANTISERA AND THE EFFECTS OF CHROMATOGRAPHY

1974 ◽  
Vol 76 (3) ◽  
pp. 597-607 ◽  
Author(s):  
John F. Hennam ◽  
William P. Collins
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Significant Difference ◽  
Assay Tube ◽  
Before And After ◽  
Process Effects ◽  
The Cost ◽  
Potential Use

ABSTRACT A method is described and evaluated for the determination of androstenedione (A) in peripheral venous plasma. The potential use of antisera to A-6-carboxymethyl thioether-bovine serum albumin and A-11α-succinyl-bovine serum albumin has been evaluated. The same plasma samples have been analysed before and after chromatography on a micro column of Sephadex LH-20. A dye, azobenzene is used to locate the fraction containing androstenedione. The results show that there is no significant difference in the values of apparent A using either antisera (overall mean 3.2 ng/ml plasma from men and 2.8 ng/ml from women). After chromatography these values are reduced by 62 and 42% respectively to 1.2 and 1.6 ng/100 ml. A new method using a mixture of ammonium and calcium sulphates is described for the separation of steroid bound to antibody. The precipitate is then resuspended and the amount of radioactivity determined, directly in the assay tube, by liquid scintillation counting. This process effects a 68 % reduction in the cost of the assay of each plasma sample.

A solid-phase enzymeimmunoassay for the determination of progesterone in bovine, porcine and ovine plasma

1995 ◽  
Vol 75 (4) ◽  
pp. 525-529 ◽  
Author(s):  
R. P. Del Vecchio ◽  
W. D. Sutherland ◽  
M. L. Connor
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Petroleum Ether ◽  
Bovine Serum ◽  
Solid Phase ◽  
Plasma Samples ◽  
Assay Sensitivity ◽  
Coefficients Of Variation ◽  
Rabbit Igg

The purpose of this project was to develop a valid quantitative enzymeimmunoassay (EIA) for progesterone in blood plasma of cattle, pigs and sheep. Rabbit anti-progesterone, mouse monoclonal anti-rabbit IgG, authentic progesterone, and acetylcholine esterase bound covalently to progesterone were the principal reagents used to develop the EIA. Ninety-six well microliter plates were coated with mouse monoclonal anti-rabbit IgG and saturated with bovine serum albumin before use. Rabbit anti-progesterone was diluted to a working dilution of 1:2.0 × 106. Standard curves were linear and ranged from 1.56 to 400 pg of progesterone per well which allowed for the measurement of 0.03125 to 8.0 ng mL−1. Assay sensitivity averaged 1.56 pg well−1. Progesterone was extracted from plasma samples with petroleum ether. Plasma samples (n = 3 or 4 from each species) with unknown amounts of progesterone that were extracted and serially diluted with EIA buffer did not deviate from parallelism with progesterone standard curves in buffer. The correlation between EIA and radioimmunoassay (RIA) measurements of progesterone in the same plasma samples was high (P < 0.0001) for all three species (r = 0.96 for bovine; r = 0.96 for porcine; r = 0.94 for ovine). The regression of EIA data on RIA data produced the following equations:[Formula: see text]The intra- and inter-assay coefficients of variation were 5.4 and 10.6% for bovine, 5.8 and 11.0% for porcine and, 6.1 and 12.3% for ovine, respectively. These data show that this EIA is a valid and reliable memod for quantitating progesterone in extracts of bovine, porcine and ovine plasma. Key words: Enzymeimmunoassay, progesterone, plasma, bovine, porcine, ovine

scholarly journals Thermally induced conformational changes and protein–protein interactions of bovine serum albumin in aqueous solution under different pH and ionic strengths as revealed by SAXS measurements

2017 ◽  
Vol 19 (26) ◽  
pp. 17143-17155 ◽  
Author(s):  
Dmitry Molodenskiy ◽  
Evgeny Shirshin ◽  
Tatiana Tikhonova ◽  
Andrey Gruzinov ◽  
Georgy Peters ◽  
...  
Keyword(s):  
Aqueous Solution ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Conformational Changes ◽  
Interaction Potential ◽  
Protein Interactions ◽  
Bovine Serum ◽  
Protein Protein Interactions ◽  
Thermally Induced ◽  
Before And After

Temperature-induced oligomerization of albumin before and after protein melting was studied using SAXS and interpreted in terms of interaction potential.

Effect of Crystals Size, Surface Area and Pore Size of Hydroxyapatite Microspheres on the Loading Ability of Bovine Serum Albumin

2011 ◽  
Vol 197-198 ◽  
pp. 17-20
Author(s):  
Jun Ming Li ◽  
Ai Juan Wang ◽  
Yu Peng Lv ◽  
Bai Ling Jiang
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Specific Surface ◽  
Pore Size ◽  
Surface Area ◽  
Specific Surface Area ◽  
Bovine Serum ◽  
Crystal Size ◽  
Adsorption Ability ◽  
Before And After

Effect of crystals size, surface area, pore size and porosity of hydroxyapatite microspheres on the loading ability of bovine serum albumin was studied in this paper. The surface morphology, specific surface area and porosity of hydroxyapatite microspheres were characterized by scanning electron microscope, specific surface area and pore size analyzer, respectively. The concentration of BSA in aqueous solutions both before and after adsorption was determined by ultraviolet-visible spectrophotometer. The results indicated that the adsorption behavior of bovine serum albumin appeared to obey the Langmuir-type isotherm model. Fast adsorption appeared at the beginning, and then decreased gradually. Hydroxyapatite microspheres calcined at 600°C had the maximum capacity, and those calcined at 800°C showed lower adsorption ability. The loading ability of hydroxyapatite microspheres depended on its crystal size, specific surface area, pore size and porosity, etc.

Human corticotropin (ACTH) radioimmunoassay with synthetic 1--24 ACTH.

1979 ◽  
Vol 25 (7) ◽  
pp. 1267-1273 ◽  
Author(s):  
P C Kao ◽  
N S Jiang ◽  
P C Carpenter
Keyword(s):  
Polyethylene Glycol ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Confidence Limit ◽  
Normal Subjects ◽  
Cross Reactivity ◽  
Normal Range ◽  
Beta Endorphin

Abstract A corticotropin antiserum was obtained from rabbits immunized with synthetic 1--24 corticotropin conjugated with bovine serum albumin. The antiserum did not cross react with synthetic alpha-melanotropin or with synthetic beta-endorphin and had a cross reactivity of 0.23% with human beta-lipotropin. We developed a radioimmunoassay with the antiserum obtained, in which we used polyethylene glycol in conjunction with a second precipitating antibody for fast (15-min) separation of antibody-bound and free corticotropin. The assay had a sensitivity of 16 ng/L and was validated on patients with various pituitary and adrenal diseases. From 103 normal subjects, the median value for corticotropin in specimens collected during the morning was 34 ng/L of plasma; the upper 95% confidence limit of the normal range was 98 ng/L.

scholarly journals The Synthesis of Wavelength-Controlled CdTe/Hydroxyapatite Composites and Their Fluorescence Enhancement by Bovine Serum Albumin

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Li Jin ◽  
Lihua Na ◽  
Xueling Cao ◽  
Fangli Zhong ◽  
Jianpo Zhang
Keyword(s):  
Quantum Dots ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Emission Intensity ◽  
Bovine Serum ◽  
Fluorescence Spectra ◽  
Fluorescence Emission ◽  
Biochemical Processes ◽  
Before And After ◽  
A Site

For the last ten years, quantum dots modified by biological materials have made it possible to study biochemical processes by means of biomedical imaging. This thesis introduced how the fluorescence CdTe quantum dots/hydroxyapatite composites were synthesized and how their structure, morphology, and fluorescence property were characterized by using TEM, XRD, EDS, UV-vis absorption spectra, and fluorescence spectra. The fluorescence spectra indicated the superb photometric characteristics of CdTe/HA composites. We also found that refluxing temperature and time had prominent effects on fluorescence wavelength and intensity of CdTe/HA composites, so the fluorescence emission wavelength of CdTe/HA composites could be controlled. In addition, the effect of BSA on the fluorescence properties of CdTe/HA composites was studied. The fluorescent emission intensity of CdTe/HA composites was enhanced directly with increasing concentrations of BSA; meanwhile, the fluorescence emission intensity of BSA dramatically decreased, which indicated that a Förster nonradiative energy transfer process occurred through the formation of chemical bonds between BSA and CdTe/HA composites. And the two-dimensional correlation (2D COS) was used to analyze the BSA solution before and after the reaction, which indicated that CdTe/HA composites have bound to a site at the surface of the molecule in the first subdomain IA. We also found that there was a linear relationship between the fluorescence intensity enhancement (F/F0) of CdTe/HA composites and the concentration of the bovine serum albumin, which might become a method for quantitative analysis of BSA in a real sample.

RADIOIMMUNOASSAY OF STEROIDS

1970 ◽  
Vol 65 (1_Suppl) ◽  
pp. S320-S331 ◽  
Author(s):  
A. R. Midgley ◽  
G. D. Niswender
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Steroid Hormones ◽  
Functional Groups ◽  
Inhibitory Activity ◽  
Bovine Serum ◽  
Major Portion ◽  
Steroid Molecule ◽  
Assay Tube

ABSTRACT A number of radioimmunoassay systems have been developed for quantitation of a variety of steroid hormones. Suitable antisera were obtained by immunizing rabbits with steroid derivatives conjugated at various positions to bovine serum albumin. The specificity of the antisera varied directly with the position of conjugation. Changes in the latter resulted in greater than 2000-fold shifts in relative inhibitory activity for a given steroid. The data indicate that conjugation should be done through a region (such as position 6) that is remote from distinguishing functional groups. The specificity data and nature of the inhibition curves suggest that the major portion of the steroid molecule is being recognized as a single immunoreactive unit. The advantages and problems associated with use of radioiodinated conjugates for labeled steroids are discussed. With conjugates, it has been possible to quantitate reliably as little as one pg of some steroids in a single assay tube.

scholarly journals Rates of pinocytic capture of simple proteins by rat yolk sacs incubated in vitro

1981 ◽  
Vol 198 (3) ◽  
pp. 581-586 ◽  
Author(s):  
G Livesey ◽  
K E Williams
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Ribonuclease A ◽  
Yolk Sac ◽  
Simple Correlation ◽  
Principal Mechanism ◽  
Molecular Charge ◽  
Before And After

The rates of pinocytic uptake of a number of small 125I-labelled simple proteins (insulin, ribonuclease A and lysozyme) by rat yolk sacs incubated in vitro were determined both before and after treating these proteins with reagents that are known to increase the rate of capture of 125I-labelled bovine serum albumin. Uptake of the untreated forms of all three proteins was extremely rapid, indicating that adsorptive pinocytosis is the principal mechanism by which yolk-sac cells capture these simple proteins, but these rates show no simple correlation with molecular charge. In contrast with albumin, the rates of uptake of treated proteins were either unchanged or lower than that of the corresponding untreated protein preparations; polymeric forms of 125I-labelled lysozyme larger than dimers were ingested at rates significantly lower than that of the monomer.

Radioimmunoassay of metanephrine and normetanephrine for diagnosis of pheochromocytoma.

1986 ◽  
Vol 32 (10) ◽  
pp. 1879-1883 ◽  
Author(s):  
K Iinuma ◽  
I Ikeda ◽  
T Ogihara ◽  
K Hashizume ◽  
K Kurata ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Mass Screening ◽  
Specific Antibody ◽  
Bovine Serum ◽  
Detection Limits ◽  
Normal Subjects

Abstract Sensitive and specific radioimmunoassays of metanephrine and normetanephrine were developed by use of 125I-labeled synephrine and specific metanephrine antibody, and 125I-labeled octopamine and specific normetanephrine antibody. Specific antibody for both metanephrine and normetanephrine was raised in rabbits by immunization with bovine serum albumin conjugated with the corresponding hapten, prepared by the method of Grota and Brown (Endocrinology 1976;98:615). The detection limits of the metanephrine and the normetanephrine radioimmunoassays were 2 and 6 pg/tube, respectively. Mean plasma metanephrine and normetanephrine values for 24 normal subjects were 62 (SD 14) and 100 (SD 40) ng/L, respectively. Mean urinary metanephrine and normetanephrine values for 22 normal subjects were 154 (SD 74) and 217 (SD 109) micrograms/day. For 14 pheochromocytoma patients, plasma metanephrine and normetanephrine values ranged from 29 to 683 and from 28 to 7850 ng/L, and urinary metanephrine and normetanephrine values were 606 to 6630 and 296 to 4800 micrograms/day, respectively. The present methods are simple and suitable for routine tests or for mass screening for pheochromocytoma.

Sign in / Sign up

Export Citation Format

Share Document