Showing papers on "Bovine serum albumin published in 1968"
TL;DR: The molecular weight-slope relation was established utilizing 17 well-characterized proteins as standards and it is possible to determine the molecular weight of a protein with an average precision of ±4%.
1,823 citations
TL;DR: Glutaraldehyde was found to react with the α-amino groups of amino acids, the N-terminal amino groups of some peptides and the sulfhydryl group of cysteine, and the phenolic and the imidazole rings of tyrosine and histidine derivatives were partially reactive.
491 citations
TL;DR: A spectrophotometric method which eliminates interference due to nucleic acid absorbance has been developed for determining protein concentration over the range of 5 to 180 μg/ml and the results obtained have been compared to those obtained with the Folin-Lowry and microbiuret methods.
429 citations
TL;DR: The similarities in the behavior of the BPA and BMA non-mercaptalbumin monomer fractions are somewhat surprising since the presence of these non-Mercapt albumin fractions in plasma albumin has been shown to be due to mixed disulfide formation between mercaptalbumin and plasma cysteine and glutathione.
357 citations
272 citations
TL;DR: Antibodies to the gastrin molecule were evoked in rabbits by immunization with synthetic human gastrin I conjugated to bovine serum albumin, and the antibodies produced were of the γG-immunoglobulin class.
254 citations
TL;DR: Agar-gel electrophoresis was used to study hyaluronate lyase and chondroitin sulfatase in the turbidity reducing unit (TRU) method.
Abstract: Microbial hyaluronidase and chondroitin sulfatase have been studied because of the possible associations between these enzymes and microbial mechanisms of pathogenicity. One of the most sensitive biological assays for mucopolysaccharide-degrading enzymes is the turbidity reducing unit (TRU) method (3). In most assays for these two enzymes, measurement of activity involves the conjugation of bovine serum fractions with nondepolymerized substrate in acetic acid. The turbidity produced by the conjugate or the lack of it can be directly related to the amount of substrate depolymerized in solution. Recently, a direct localization and visualization technique to study hyaluronate lyase by agar-gel electrophoresis was reported (1). This technique was modified as a cultivable screening plate method for bacteria. The basic medium consisted of Brain Heart Infusion broth (BBL) prepared to make 100 ml, to which was added 1 g of Noble agar (Difco). The medium was autoclaved at 121 C for 15 min and cooled to 46 C. Aqueous solutions of 2 mg of umbilical sodium hyaluronidate (Sigma Chemical Co., St. Louis, Mo.) per ml, 4 mg of bovine nasal chondroitin sulfate (Pentex, Inc., Kankakee, Ill.) per ml, and 5% bovine albumin fraction V (Signa) were sterilized by filtration with 0.20-,um Nalgene filter units (Nalge Co., Inc., Rochester, N.Y.). Each substrate was added to the cooled media to give final concentrations of 400 ,Ag/ml. The bovine albumin fraction-V was then added with constant stirring to give a final concentration of 1%. The agar was poured to a depth of 3 to 4 mm. The finalpH of each medium was 6.8 0.1. After solidification, plates were tempered at 4 C to provide a firm surface for streaking or swabbing. Pure or mixed cultures
198 citations
TL;DR: It is reported that aspirin acetylates a variety of body constituents in vitro, including HSA, serum albumin, and other biological materials.
Abstract: HUMAN serum albumin (HSA) is permanently altered after exposure to therapeutic concentrations of acetylsalicylic acid (ASA, aspirin) either in vitro or in vivo1–3. Recently we showed that this structural alteration was the result of the acetylation of the HSA molecule by aspirin4,5. Because the acetylation of HSA by aspirin could occur by aminolysis resulting in an N-acetyl-HSA derivative, we decided to determine whether aspirin could acetylate other biological materials. We now report that aspirin acetylates a variety of body constituents in vitro.
174 citations
TL;DR: The Standards Committee of the AACC presents a discussion of the nature of total serum protein and the problems associated with its determination, and proposes that protein be defined as polypeptide material, and that a reference preparation be promulgated for interlaboratory use.
Abstract: The Standards Committee of the AACC presents a discussion of the nature of total serum protein and the problems associated with its determination. Proposals are made that protein be defined as polypeptide material, and that a reference preparation be promulgated for interlaboratory use. The reference material suggested is bovine serum albumin, produced to rigid specifications and distributed as a stable 7% (w/v) solution, the concentration of which has been established by careful dry weight assay. Comments of readers are invited.
140 citations
TL;DR: It was found that both antibody-forming cells and their precursors were present in the denser region of the gradient when spleen cell suspensions were taken from unimmunized mice, and both antibodies and precursor floated to the top in cell suspensions from mice sacrificed 1, 2, or 3 days after antigen injection.
Abstract: Cell suspensions from the spleens of normal mice or mice injected with sheep erythrocytes were separated on a discontinous bovine serum albumin density gradient. Four bands or subpopulations were obtained and were assayed for antibody-forming cells, and for antigen-sensitive precursor cells. The antibody-forming cells were assayed by the hemolytic plaque assay and the antigen-sensitive precursors were assayed by the number of plaque-forming cells which developed after 3 or 5 day's culture with antigen. It was found that both antibody-forming cells and their precursors were present in the denser region of the gradient when spleen cell suspensions were taken from unimmunized mice. In contrast, both antibody-forming cells and precursors floated to the top in cell suspensions from mice sacrificed 1, 2, or 3 days after antigen injection. The change in density was detectable as early as 12 hr and was complete by 18 hr. The cell which changed in density was specific for the antigen that brought about that change. The significance of these findings is discussed.
135 citations
TL;DR: In this paper, reaction rates with acrylic acid derivatives were determined for the NH2 and SH groups in mercaptoethanol and in the reduced wheat gluten and bovine serum albumin.
TL;DR: It was found that female mice developed a stronger and longer lasting immune response and thatfemale mice were more responsive to small doses of antigen.
Abstract: SummaryThe immune response to a protein antigen (BSA, bovine serum albumin) was compared in male and female mice. It was found that female mice developed a stronger and longer lasting immune response and that female mice were more responsive to small doses of antigen. An explanation based on the effect estrogens have on phagocytosis is discussed.
TL;DR: With these preparations, it has now been possible to show a substrate-dependent and stoichiometric decarboxylation of a--ketoglutarate coupled to the hydroxylations of peptidyl proline residues.
Abstract: The collagen proline hyd.roxylase system (also referred to as protocollagen hydroxylase), which converts selected proline residues to hydroxyproline in polypeptide precursors of collagen, has now been purified from a variety of animal tissues.1-4 M\/Iost purified preparations have an absolute requiremenlt for Fe++ and for a reducing agent, which can be ascorbic acid, other enediols, or tetrahydropteridines. An additional requirement discovered by Hutton et al.1' is a-ketoglutarate. This keto acid cannot be replaced by pyruvate, oxaloacetate, or related compounds and is an absolute requirement for proline hydroxylation. Removal of the keto acid by incubation with crystalline glutamic dehydrogenase, NH4+, and reduced nicotinamide-adeniine dinucleotide (NADH) leads to total inactivation of crude homogenates; activity is restored by the addition of aketoglutarate. Earlier attempts in this laboratory to determine whether a-ketoglutarate was consumed during the course of the hydroxylation reaction were inconclusive because the chick-embryo enzyme preparations used contained other enzymes that metabolized a-ketogluftarate.1 More recently, we have been able to obtain preparations from fetal, rat; skin that are far more active with respect to proline hydroxylation and that do not contain competing enzymes. With these preparations, it has now been possible to show a substrate-dependent and stoichiometric decarboxylation of a--ketoglutarate coupled to the hydroxylation of peptidyl proline residues. Ml/aterials and M11ethods.-Both H(3,4-Hl3-Pro-Gly-Pro),OH (mol wt 4100; spee. act. 0.9 mec/nmg) and poly-L-proline 11 (mol wt 9000) were gifts from Drs. J. Kurtz and A. 13erger, Weizmanari I nstitute of S ciences, Rehovoth, Israel. We obtaiiied a-ketoglutarate5-C14, (9.33 me/mnmole) from Nuelear-Chicago Corp., and a-ketoglutarate-U-C14 (3.2 me/mmole) an-d a-ketoglutarate-1-C14 (11 me/mmole) from Calbiochem, Inc. These were diluted with nonradioactive a-ketogluitarate before use. Crystalline bovine liver L-glutamic acid dehydrogenase) crystalline bovine liver catalase, and NADH were all products of Sigma Chemical Company. NCS reagent (for trapping C02) was obtained from Nuclear-Chicago Corp., and crystalline bovine serum albumin from Pentex Corp. Collagen proline hydroxylase was prepared from skins of newborn rats. Approximately 300 gm of tissue were homnogenized in 2 vol of 0.25 M. sucrose containing 0.01 mM ethylenediaini,1etet-aacetate (E})TA) aid O.l OJmM dithiothreitol, and the homogenate was centrifuged at 27,000 X g. All solutions used in subsequent purificatioin contained the same coneentrations of ditliothieitol. TFwo vol of saturated ammonium sulfate at pH 7.0 were added to the supelrnatant fraetion, and after centrifugation, the precipitate was dissolved in sodium cacodylate buffer, pH 7.0, and treated with 0.64 mg streptomycin sulfate per mg protein to remove nucleic acids. After centrifugation for 1 hr at 68,000 X g, the supernatanit fluid was treated with nieutral ammonium sulfate, and the fraction that precipitated between 17%( and 39% saturation (12 and 27 gm/100 ml) was dissolved in 0.05 M cacodylate, pH 7.0, containing 0.2 11f NaCl. After dialysis against 4 liters of the same solution, the en-zyme (800 mg of protein) was charged onto an O-(diethyl-
TL;DR: The tetrapeptide l-aspartyl- l-threonyl-l-histidyl-l -lysine was shown by titration, absorbance, and circular dichroism measurements to form an equimolar complex with copper(II) that has substantially the properties of the complex of peptide (1–24) containing 1 eq of copper( II) ion.
TL;DR: It is suggested that protein bound glutathione may represent a reservoir of physiological radiation protector and be of significance or the radio protective action of cysteamine.
TL;DR: A regimen of small doses of the complexes elicited high titers of both types of antibodies in eight out of eight rabbits, indicating a potent immunogen for the elicitation of both anti-BSA and anti-ACTH antibodies.
Abstract: SummaryA method is described for the uniform elicitation of antibodies in rabbits to ACTH. Porcine ACTH and bovine serum albumin have been covalently cross linked with glutaraldehyde. This complex is non-steroidogenic and a potent immunogen for the elicitation of both anti-BSA and anti-ACTH antibodies. A regimen of small doses of the complexes elicited high titers of both types of antibodies in eight out of eight rabbits. These titers are still present 6 months after the last injection of antigen.
TL;DR: The results indicate that in rat embryos the cerebral blood vessels are impermeable to albumin at least as early as the fifteenth day after fertilization.
Abstract: A study was made on the permeability of cerebral blood vessels to albumin during development. Fluorescent labeled bovine serum albumin was injected into a tail vein of newborn, young and adult rats and into the umbilical artery of embryos from the fifteenth to the twenty-first day of pregnancy. The distribution of the tracer was ascertained by means of fluorescence microscopy. In the brains of the embryos and postnatal rats the fluorescent albumin was strictly confined to the lumen of the blood vessels, while considerable extravascular passage was observed in subcutaneous tissue. The results indicate that in rat embryos the cerebral blood vessels are impermeable to albumin at least as early as the fifteenth day after fertilization.
TL;DR: In vitro studies demonstrated that a major part of the antibodies produced by rabbits with chronic nephritis lacked precipitating properties, and it is suggested that, in addition to quantity, quality of antibody plays an important role in the development of chronic serum sickness.
Abstract: Three of 16 rabbits injected (intravenously) daily with crystalline bovine serum albumin (BSA) for periods in excess of 10 wk developed chronic glomerulonephritis. In vivo, animals with chronic proteinuria formed variable quantities of soluble complex after injection of antigen while animals without proteinuria exhibited rapid removal of the injected BSA. In vitro studies demonstrated that a major part of the antibodies produced by rabbits with chronic nephritis lacked precipitating properties. Interpretations of these observations were presented in the discussion. It is suggested that, in addition to quantity, quality of antibody plays an important role in the development of chronic serum sickness. Complexes formed with nonprecipitating antibody, which are less rapidly removed from circulation, would have a greater opportunity to deposit in glomeruli and induce inflammation.
Journal Article•
TL;DR: The stimulatory effect of antigen–antibody–complement complexes on cultured normal human peripheral blood lymphocytes was studied and the most likely explanation of this stimulation is injury to lymphocyte membranes, possibly from a non-specific attachment of immune complexes to them.
Abstract: The stimulatory effect of antigen–antibody–complement complexes on cultured normal human peripheral blood lymphocytes was studied. The donors of the cells had not been sensitized to the antigens used. Two antigens were used: flagellar antigen of Salmonella paratyphi B (SPB) and bovine serum albumin (BSA), with their respective antibodies prepared in rabbits. The addition of such antigen–antibody aggregates to the cultures stimulated the lymphocytes as determined by morphological changes and increased uptake of [14C]thymidine into DNA. Peak of stimulation was observed after 5–6 days of culture incubation. The stimulation appeared to be complement dependent. The lymphocytes showed no response either to the antigen alone or to anti-SPB. When BSA–anti-BSA was centrifuged, most of the stimulatory activity was found in the supernate. The most likely explanation of this stimulation is injury to lymphocyte membranes, possibly from a non-specific attachment of immune complexes to them. A similar mechanism of membrane injury may underlie reactions to all non-specific stimulants, and possibly also to specific antigens to which the cell donor is sensitized.
TL;DR: The behavior of lysozyme in aqueous urea and acetamide is consistent with the following conclusions: like serum albumin, the extent of solute-protein interaction is paralleled by some degree of protein perturbation, the latter reflected by ly sozyme in change of intrinsic viscosity or disulfide bond stability (or both).
TL;DR: The partitioning of several naphthylamine sulphonate derivatives between water and solutions of dodecylamine in hexane andbetween water and detergent micelles was studied as an aid to understanding the binding of these dyes to bovine serum albumin.
TL;DR: It is shown that the transfer of thymus cells alone to irradiated recipient mice would not restore their antibody response to sheep erythrocyte antigens, but that these cells would act in cooperation with bone marrow cells when a mixture of these cell types was transferred.
Abstract: IT has been observed consistently that the immune responsiveness of neonatally thymectomized mice to various antigens, including bovine serum albumin (BSA), can be restored by injection of sufficient dissociated thymus cells1,2. This is also true for mice thymectomized in adult life, lethally irradiated and injected with bone marrow (my unpublished data). Claman, Chaperon and Triplett3 have shown that the transfer of thymus cells alone to irradiated recipient mice would not restore their antibody response to sheep erythrocyte antigens, but that these cells would act in cooperation with bone marrow cells when a mixture of these cell types was transferred.
TL;DR: The results suggest that testosterone binding may alter bovine serum albumin conformation and that the measured binding capacity of this protein for testosterone is dependent on the protein concentration.
TL;DR: Adversity of antibodies and their subsequent elution at pH 6·–6·8, using ions such as thiocyanate as the eluent, consistently result in recovery of from one-half to three-fourths of the active, precipitable antibody present in the original solutions.
Journal Article•
TL;DR: The amount of protein in the extravascular fluid spaces after equilibration, as measured by the CL/CP, correlated exponentially with the effective hydrodynamic diffusion radius of the protein.
Abstract: Plasma proteins varying in molecular weight from 50,000 to 950,000 were radiolabeled and injected into rabbits and the lymph to plasma concentration ratios (CL/CP) of the various proteins were measured 3 days following injection. The amount of protein in the extravascular fluid spaces after equilibration, as measured by the CL/CP, correlated exponentially with the effective hydrodynamic diffusion radius of the protein. An exponential relationship also was observed between CL/CP and molecular weight for globular proteins, whereas fibrinogen, which is a spindle-shaped protein, showed only a correlation with the effective hydrodynamic diffusion radius and not with the molecular weight. No correlation between the intra- and extravascular distribution and half-lives of the various homologous and heterologous proteins was observed. The immunoglobulins of similar molecular size, but different chemical structure, as human γA-monomer, human γG and rabbit γG, showed a similar lymph to plasma concentration ratio following equilibration.
Following intravenous injection, a larger percentage of bovine serum albumin (BSA) complexed with Fab fragments from purified anti-BSA remained in the intravascular fluid spaces than of BSA alone after complete equilibration.
Similar to the CL/CP, the calculated extravascular lymph space occupied by the various proteins correlated exponentially with the effective hydrodynamic diffusion radius of the protein.
The importance of the in vivo distribution and the behavior of protein antigens in relation to immunologic phenomena is discussed. With a given blood concentration, the concentration of serum protein antigens in the extravascular fluid space varies considerably with antigens of different molecular sizes.
TL;DR: The binding of methyl orange by bovine serum albumin is suppressed by urea, but the inhibition is largely reversible if urea is removed either by dialysis (fast or gradient) or by dilution.
TL;DR: The transfer of imunOGLOBULins and serum albumin from blood into milk of lactating EWes and serum alcohol from blood Into Milk of LACTATING EWES is described.
Abstract: THE TRANSFER OF [ 131 I]-LABELLED IMMUNOGLOBULINS AND SERUM ALBUMIN FROM BLOOD INTO MILK OF LACTATING EWES