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Showing papers on "Bovine serum albumin published in 1972"


Journal ArticleDOI
TL;DR: Under the new conditions there is direct proportionality between absorbance at 650 nm and weight of protein within the range 15–110 μg.

5,177 citations


Journal ArticleDOI
TL;DR: A modification of the Lowry procedure for the rapid and reliable analysis of proteolipid protein involves the addition of sodium dodecyl sulfate to dissolve the lipid-containing protein samples or to maintain the water-soluble apoprotein in solution.

555 citations



Journal ArticleDOI
TL;DR: A double antibody radioimmunoassay for TRH has been developed in which as little as 0.1 ng unlabeled TRH can be detected and a variety of TRH analogues were tested and all had little reactivity in this immunoassays.
Abstract: Synthetic thyrotropin releasing hormone (L-pyroglutamyl-L-histidyl-L-prolineamide) (TRH) was coupled to bovine serum albumin (BSA) with bis-diazotized benzidine (BDB). Recovery after dialysis of 14C-TRH included in the reaction mixture indicated that the product contained 37.5 μg TRH/mg albumin. The TRH-BDBBSA conjugate in doses calculated to contain 200 and 1000 ng TRH was biologically inactive whereas 20 ng TRH significantly increased rat serum TSH in vivo. Immunization of rabbits with the conjugate yielded significant quantities of anti- TRH antibody which was detected by reaction of diluted antiserum with 125I-TRH. Using 125I-TRH a double antibody radioimmunoassay for TRH has been developed in which as little as 0.1 ng unlabeled TRH can be detected. A variety of TRH analogues were tested and all had little reactivity in this immunoassay. The most potent of these was L-pyroglutamyl-D-histidyl-L-prolineamide which had 1.08% of the activity of TRH. Addition of the amino acid constituents of TRH, various ...

236 citations


Journal ArticleDOI
TL;DR: The greater degree of fluorescence enhancement, the greater stability ofretinol in RBP and PA-RBP to spontaneous degradation at room temperature, the relative resistance to extraction of the bound retinol by ether, and the complete unavailability of the retinols inRBP andPA-R BP for oxidation by liver alcohol dehydrogenase were interpreted as indicating that retinOL is bound with greater affinity to the natural carrier protein than it is to bovine serum albumin or

234 citations


Journal ArticleDOI
TL;DR: The effectiveness with which amino acids compete with human serum albumin for zinc was shown to depend upon the relevant metal complex stability constants, and it is indicated that cysteine and histidine are the only important amino acid ligands of zinc in plasma.

216 citations


Journal ArticleDOI
TL;DR: This assay procedure, which avoids chromatography and uses only one organic solvent extraction had blank values which approached zero, and introduction of an additional thin layer chromatographic step was associated with ...
Abstract: A globulin fraction rich in bound testosterone was separated from serum by ammonium sulfate precipitation. Testosterone and related steroids were quantitatively extracted from this fraction with a benzene petroleum ether mixture and measured by radioimmunoassay using testosterone-3-tyrosine methyl ester-125I and a specific antiserum elicited against testosterone-3-oxime bovine serum albumin. This assay procedure, which avoids chromatography and uses only one organic solvent extraction had blank values which approached zero. The within and between assay error, measured as the coefficient of variation was 7.8% (n = 64) and 8.2% (n = 15), respectively. The minimum amount of hormone per sample which can be measured with good reliability is 0.17 ng. This provides sufficient hormone to account for losses during the procedure, and to allow quadruplicate samples for radioimmunoassay and an aliquot for determining recovery. Although introduction of an additional thin layer chromatographic step was associated with ...

211 citations


Journal ArticleDOI
18 Aug 1972-Science
TL;DR: It is suggested that local immunization interferes specifically with the intestinal uptake of macromolecular antigens in rats immunized with peroxidase and bovine serum albumin.
Abstract: Animals were orally immunized with horseradish peroxidase and bovine serum albumin, and absorption of these antigens was studied. In comparison with controls, a consistent and significant decrease in peroxidase uptake was noted in both germ-free and conventional rats immunized with peroxidase; a similar decrease in serum albumin uptake was also noted in animals immunized with serum albumin. There was no difference in the uptake of an unrelated macromolecule. These observations suggest that local immunization interferes specifically with the intestinal uptake of macromolecular antigens.

204 citations


Journal ArticleDOI
TL;DR: Results indicate that lipid A contains the endotoxic principle of bacterial lipopolysaccharide, which is generally less active than endotoxic lipopoly Saccharides.
Abstract: Lipid A preparations from the lipopolysaccharides of four Salmonella minnesota R mutants and one Escherichia coli 0100 R mutant were assayed as soluble complexes with bovine serum albumin for lethality in mice, pyrogenicity in rabbits and complement inactivation. Although generally less active than endotoxic lipopolysaccharides, these lipid · albumin complexes nevertheless exhibited strong biological activity. These results indicate that lipid A contains the endotoxic principle of bacterial lipopolysaccharides.

200 citations


Journal ArticleDOI
TL;DR: An antibody obtained by immunizing a ewe with 11α-hydroxy-progesterone-hemisuccinate conjugated to bovine serum albumin was used for the assay of progesterone in crude plasma extracts and accuracy was confirmed by the recovery of known amounts of progestersone added to plasma.
Abstract: A specific progesterone antibody was obtained by immunizing a ewe with 11α-hydroxy-progesterone-hemisuccinate conjugated to bovine serum albumin. This antibody was used for the assay of progesterone in crude plasma extracts. The sensitivity of the assay was 0.05 ng. The accuracy was confirmed by the recovery of known amounts of progesterone added to plasma. The inter- and intra-assay coefficient of variation was 8%. Specificity was confirmed by a) checking cross reactivity of various steroids; b) failure of other steroids added to plasma to influence the accurate measurement of progesterone, and c) finding comparable progesterone levels in plasma extracts with and without chromatography. Mean values of progesterone (ng/ml) were 0.26 ± 0.08 (sd) in males and postmenopausal females; 0.50 ± 0.44 in preovulatory females; and 12.1 ± 4.81 in postovulatory females.

196 citations


Journal ArticleDOI
01 Mar 1972-Steroids
TL;DR: It is concluded that the site of attachment of steroid haptens to the peptide carrier importantly affects the specificity of the antibodies produced.

Journal ArticleDOI
TL;DR: The cytoplasm-to-nucleus transfer of proteins was limited by their molecular size, and small proteins, neutrally and positively charged, showed rapid nuclear incorporation.

Journal ArticleDOI
TL;DR: A radioimmunoassay developed using dextran-coated charcoal to separate the free from antibody bound PGA1-3H is sufficiently sensitive, precise, and rapid to allow the routine estimation of the PGAs in biological samples.

Journal Article
TL;DR: The quantitative distribution of immunoglobulins and albumin were measured in serum and nasal, lacrimal, salivary and vaginal secretions of four heifers for 9- to 12-week periods and with two exceptions a significant positive correlation existed between IgG andalbumin in every secretion.
Abstract: Immunoglobulins in bovine serum and in lacteal, salivary, nasal and vaginal secretions have been characterized on the basis of antigenic distinctions, ion exchange and gel filtration properties, and ultracentrifugal analysis IgG 1 , IgG 2 , IgM and IgA have been clearly distinguished and the presence of a secretory IgA system analogous to that in other mammalian species has been confirmed All detectable bovine serum IgA has a molecular weight consonant with its existence in dimeric form The quantitative distribution of immunoglobulins and albumin were measured in serum and nasal, lacrimal, salivary and vaginal secretions of four heifers for 9- to 12-week periods IgA is a minor serum component but is the predominant immunoglobulin in nasal secretions, tears and saliva; in vaginal mucus its relative concentration is lower than IgG but still much higher proportionally than in serum Marked weekly fluctuations occurred in each secretion in IgA/total protein ratios IgG 1 was consistently the predominant IgG subclass in each secretion although the serum relationship varied among animals With two exceptions a significant positive correlation existed between IgG and albumin in every secretion IgM levels in secretions were generally low but in two parenterally immunized heifers mean IgM concentrations in nasal secretions exceeded those of IgG Local synthesis or selective transport must account for virtually all of the IgA as well as a portion of IgG 1 and, possibly, IgM in these secretions Serum components derived by transudation account for a much higher proportion of total protein in vaginal mucus than in nasal, lacrimal, or salivary secretions

Journal ArticleDOI
01 Dec 1972-Steroids
TL;DR: A radioimmunoassay procedure for the determination of testosterone in plasma is described, which makes use of the new anti-testosterone serum and preliminary results suggest that it can be applied to ether extracts from human sera without Chromatographic purification.

Journal ArticleDOI
TL;DR: Studies with undefatted and defatted monomers indicate that one of the primary fatty acid binding sites is also located in this part of the crevice and that oxidation of the cysteinyl sulfhydryl group alters the binding of fatty acid at this site.

Journal ArticleDOI
TL;DR: It is suggested that B and T cells recognize different epitopes on the BSA molecule, only those recognized by B cells being affected by the methylation procedure.
Abstract: Immune cells induced by bovine serum albumin (BSA) and its methylated derivative (MBSA) have been compared in a cooperative cell transfer system for their content of BSA-specific antibody-forming cell precursors (AFCP, B) and BSA-specific helper (T) cells. When MBSA immune cells were transferred together with hapten-primed cells into recipient mice which were stimulated by a hapten-BSA conjugate, their cooperative secondary anti-hapten response was as good as in case of transferred BSA immune cells. Their secondary anti-BSA response, however, was markedly reduced (reduction factor > 30). Hapten-MBSA conjugates had the same capacity to react with BSA-specific helper cells in the cooperative secondary anti-hapten response as hapten-BSA conjugates but had a reduced ability to react with BSA-specific AFCP cells. In spite of the pronounced reduction of the B cell response, MBSA had the same threshold dose as BSA for activating BSA-specific T cells. These data suggest that B and T cells recognize different epitopes on the BSA molecule, only those recognized by B cells being affected by the methylation procedure.

Journal ArticleDOI
TL;DR: The effectiveness of the copper IUD may arise in 2 general ways: 1) the scission of disulfide bonds resulting in a modification of S-S containing proteins and mucus causing precipitation of albumins which may change the internal wall in such a way as to prevent implantation and 2) the presence of oxygen.

Journal ArticleDOI
TL;DR: In this paper, the authors showed that the effect of albumin when ACD blood is frozen was demonstrated, and that albumin increased freezing damage at faster cooling rates than other non-penetrating agents.

Journal ArticleDOI
TL;DR: Kinetic studies showed that the two p-nitrophenyl substrates tested compete for the active sites on both enzymes, and enzyme structure could not be deduced by fitting with calculated curves.

Journal ArticleDOI
09 Jun 1972-Science
TL;DR: Morphine-3-hemisuccinate was synthesized by reaction of morphine with succinic anhydride, and the degree of conjugation was determined by base hydrolysis of the conjugate, extraction, and measurement of free morphine.
Abstract: Morphine-3-hemisuccinate was synthesized by reaction of morphine with succinic anhydride. This compound was conjugated to bovine serum albumin by the mixed anhydride method, and the degree of conjugation was determined by base hydrolysis of the conjugate, extraction, and measurement of free morphine. An average of 6.5 molecules of morphine were conjugated to each molecule of protein. Eleven rabbits immunized with varying doses of the conjugate were producing antibody 8 weeks later, as determined by a modification of the ammonium sulfate method, which measures primary binding of antigen by antibody.

Journal ArticleDOI
TL;DR: Analysis of albumin which has reacted with p-nitrophenyl acetate indicate that two tyrosyl hydroxyl groups on the protein are rapidly acetylated by the ester.

Journal ArticleDOI
TL;DR: Raman spectra of aqueous bovine serum albumin and β-lactoglobulin were obtained with 4880 A radiation of an argonion laser as mentioned in this paper.

Journal ArticleDOI
TL;DR: Catheptic activity was found to be localized in the lysosomal fraction of skeletal muscle of winter flounder and the enzyme hydrolyzed hemoglobin, globin, bovine serum albumin and endogenous proteins.
Abstract: Catheptic activity was found to be localized in the lysosomal fraction of skeletal muscle of winter flounder (Pseudopleuronectes americanus). The enzyme hydrolyzed hemoglobin, globin, bovine serum albumin and endogenous proteins. The hemoglobin splitting activity of the enzyme under different conditions was determined by gel electrophoresis. An optimum pH of 4.0 was found for both the hemoglobin breakdown and autolytic activity on the endogenous proteins. The enzyme was highly thermostable, dialysis had no effect on its activity, and the enzyme was inhibited by sodium chloride, cysteine, ATP, NAD and phenyl methyl sulfonyl fluoride. Its molecular weight was found to be about 32,000 using sucrose gradient centrifugation.

Journal ArticleDOI
TL;DR: Tryptophan fluorescence spectra of the native lipoprotein and of its delipidated form have maximum fluorescence wave lengths at 328 and 338 nm, respectively, an indication that on delipidation the environment of tryptophan residues becomes more polar.

Journal Article
TL;DR: CPS-K could induce the antibody response of mice to syngeneic eyeball extracts which are normally non-immunogenic and was also much stronger than that of lipopolysaccharide of Escherichia coli and zymosan.
Abstract: The capsular polysaccharide of Klebsiella pneumoniae (CPS-K) was shown to enhance markedly antibody response of mice to various antigens. The primary antibody response to bovine serum albumin (BSA) occurred more rapidly after an injection of BSA mixed with CPS-K than after injection of the same amount of BSA in Freund9s complete adjuvant (CFA), although the peak antibody level in the serum after the primary injection became higher eventually in the latter mice. In secondary and tertiary responses, much more pronounced antibody response (occasionally more than 30 mg of antibody protein per ml) was induced in mice injected first with BSA mixed with CPS-K than in those injected originally with BSA in CFA. The adjuvant action of CPS-K was also much stronger than that of lipopolysaccharide of Escherichia coli and zymosan. CPS-K could induce the antibody response of mice to syngeneic eyeball extracts which are normally non-immunogenic.

Journal ArticleDOI
TL;DR: The occurrence of these immunoglobulin units helps to explain the complicated immunoelectrophoretic patterns often observed in bovine body fluids and also the multiple ring phenomena obtained during radial diffusion assay for Immunoglobulins in the cow.

Journal ArticleDOI
TL;DR: It is concluded that the amounts of TeBG present in serum regulate the supply of the hormone to the target tissue and thus control its biological action.
Abstract: The influence of serum on the uptake, conversion and action of dihydrotestosterone in relation to the sex steroid binding protein, TeBG, has been investigated in rat ventral prostates in organ culture. The organs were incubated with (1,2-3H)dihydrotestosterone in: (1) serum-free medium, (2) horse serum, foetal and newborn bovine serum of (3) human male and human pregnancy serum. With all sera the uptake of dihydrotestosterone fell with rising serum concentration, at first steeply and then more gradually. At the same concentration, the uptake was significantly lower in explants incubated with human pregnancy serum than in those kept with human male serum. The conversion of dihydrotestosterone to androstanediol followed the same pattern and less androstanediol was formed in the presence of pregnancy serum. Since pregnancy serum contains higher amounts of TeBG than male serum, the lowered uptake suggests that only the free hormone was available to the target organ. Addition of unlabelled dihydrotestosterone resulted in a higher uptake than that measured in explants incubated with the labelled steroid only. The effect of the human sera on uptake and conversion was correlated with the androgenic activity of dihydrotestosterone applied at physiological concentrations and expressed as the percentage of secretory columnar cells present. The degree of maintainance closely corresponded to the uptake of the hormone. In serum-free medium, the number of columnar cells approached the values found in vivo, with male serum their number, though reduced, was still substantial, with pregnancy serum it was extremely low. It is concluded that the amounts of TeBG present in serum regulate the supply of the hormone to the target tissue and thus control its biological action.

Journal ArticleDOI
01 Dec 1972-Steroids
TL;DR: A radioimmunoassay for plasma corticosterone has been developed that was practical and reliable and produced results similar to that obtained by other methods.

Journal ArticleDOI
TL;DR: Phospholipase A2 activity in lung supernatant was considerably higher than that obtained in a liver preparation and was optimal at pH 6.5 and was strongly inhibited by sodium deoxycholate.