The Choline and Serum Protein Requirements of Mouse Fibroblast Cells (Strain LS) in Culture

1969 ◽  
Vol 5 (1) ◽  
pp. 135-142
Author(s):  
J. R. BIRCH ◽  
S. J. PIRT
Keyword(s):  
Bovine Serum Albumin ◽  
Population Density ◽  
Serum Albumin ◽  
Serum Protein ◽  
Cell Population ◽  
Bovine Serum ◽  
Choline Chloride ◽  
Mouse Fibroblast ◽  
Population Doubling ◽  
Maximum Cell

The maximum cell population density of mouse fibroblast (strain LS) cells growing in static suspension culture was found to be directly proportional to the dialysed calf serum concentration. This was due to choline limitation and the fact that serum protein was a major source of choline. The growth yield (Y) was 3.2 x 105 cells/µg choline chloride. Studies on the role of serum in the presence of excess choline showed the following. When protein was omitted from the medium, cell death occurred. Whole serum protein could be replaced by either (1) bovine serum albumin fraction V, or (2) crystalline bovine serum albumin + sodium pyruvate and α-ketoglutarate, or (3) polyvinylpyrrolidone + methylcellulose + pyruvate and α-ketoglutarate. The population doubling time was 24 h in the presence of whole serum protein and increased considerably with the substitutes (1-3). The increase in maximum cell population density (without medium changes) exceeded 2.9 x 106 cells/ml with either whole serum protein or substitutes (1) and (2). With serum substitute (3) the maximum increase in population density was reduced to 1.6 x 106 cells/ml.

Glycylglycylglycine as a Synthetic Standard for Serum Protein Determination

1974 ◽  
Vol 20 (1) ◽  
pp. 70-73
Author(s):  
Bernard Klein
Keyword(s):  
Human Serum Albumin ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Serum Protein ◽  
Bovine Serum ◽  
Molar Absorptivity ◽  
Protein Determination ◽  
Synthetic Standard ◽  
Mean Differences

Abstract Glycylglycylglycine was investigated as a reference standard for use in serum protein measurement by the biuret reaction. The tripeptide-biuret solution has a molar absorptivity of 96 at 565 nm, and absorbances at both 550 nm and 565 nm are proportional to concentration. By a manual reference procedure, the 550-nm absorbance of 1.0 g of tripeptide was equivalent to that given by 1.72 ± 0.03 g of human serum albumin or 1.43 ± 0.03 g of bovine serum albumin. By the Technicon N14b automated procedure, the absorbance of 1.0 g of tripeptide at 550 nm was equivalent to that of 1.81 ± 0.02 g of human serum albumin or 1.89 ± 0.03 g of bovine serum albumin. Results for serum protein analyses over the range 4.0 to 9.0 g/dl, when tripeptide or serum albumin was used to prepare calibration curves, showed mean differences of 0.15 g/dl in the manual mode and 0.08 g/dl in the automated mode.

HISTAMINE-LIBERATING ACTIVITY OF SERUM PROTEIN EXPOSED TO DEAE-CELLULOSE

1967 ◽  
Vol 45 (3) ◽  
pp. 479-489
Author(s):  
M. Levy ◽  
J. Q. Bliss
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Serum Protein ◽  
Bovine Serum ◽  
Active Material ◽  
Local Treatment ◽  
Deae Cellulose ◽  
Autologous Serum ◽  
Endogenous Histamine ◽  
Simple Exposure

Dog serum protein fractions prepared by chromatography on DEAE-cellulose produced an increase in local small-vessel permeability with distinct wheal formation when injected intracutaneously into the animal from which the whole serum was obtained. This permeability activity also appeared when the serum was left unseparated by simple passage through a DEAE-cellulose column in terminal buffer. Bovine serum albumin also developed permeability activity when treated this way. Neither whole autologous serum nor bovine serum albumin left untreated, showed any permeability activity. The whealing responses to active material were always reduced or abolished (a) after the test animal was given 2 mg/kg of the antihistamine mepyramine, and (b) in areas of skin depleted of their endogenous histamine by local treatment with compound 48/80. It is suggested that simple exposure of some protein molecules to the DEAE grouping converts them into histamine liberators.

Standards for Total Serum Protein Assays— A Collaborative Study

1975 ◽  
Vol 21 (8) ◽  
pp. 1159-1166 ◽  
Author(s):  
Basil T Doumas
Keyword(s):  
Human Serum Albumin ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Serum Protein ◽  
Bovine Serum ◽  
Total Serum Protein ◽  
Total Serum ◽  
Final Reaction ◽  
Protein Assays

Abstract We have studied the standardization of total serum protein assay with the biuret reaction. Standard solutions were prepared from lyophilized preparations of human serum albumin and bovine serum albumin, with corrections made for volatile material and ash contents. These solutions and a solution of crystalline albumin standard were analyzed with a new stable biuret reagent, to establish absorptivity values (values for the absorbance of a 1 g/liter final reaction mixture). The mean values obtained were 0.302, 0.292, and 0.290 for human serum albumin, bovine serum albumin, and the crystalline albumin, respectively. We believe that the established absorptivity value will improve the accuracy of serum protein determinations. We studied the linearity of the relation between color produced and protein concentration, with use of the solutions described above and a serum pool. The color adheres to Beer's law up to the highest concentration tested: 3 g/liter for HSA and BSA, and 2.8 g/liter for serum in the final reaction mixture. The new biuret reagent has been stable for one year at room temperature. We recommend the use of bovine serum albumin as a primary standard for serum protein assays. It is inexpensive, easily available, and exhibits the best linearity in the biuret reaction.

Research on the Interaction between Pheophorbide and Bovine Serum Albumin

2013 ◽  
Vol 749 ◽  
pp. 471-476 ◽  
Author(s):  
Yong Ye ◽  
Xue Lan Chen ◽  
Ya Guo
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Serum Protein ◽  
Bovine Serum ◽  
Fluorescence Spectra ◽  
Synchronous Fluorescence ◽  
Tryptophan Residue ◽  
Hydrophobic Force ◽  
Synchronous Fluorescence Spectra ◽  
Spontaneous Interaction

The interaction between natural pheophorbide (a superior photosensitizer) and bovine serum albumin (BSA) in physiological condition is investigated by means of UV-Vis, fluorescence and synchronous fluorescence spectra so as to provide the basis for clinical use. Natural pheophorbide was isolated from silkworm excrement. BSA in pH 7.4 Tris buffer mixed with different concentration of pheophorbide was kept at certain temperature for 3 h or under illumination by laser at 630 nm for 20 min. UV-Vis absorption of BSA was enhanced and its fluorescence was quenched by pheophorbide. Illumination of laser at 630 nm intensified the quenching. The mechanism is deemed as mainly static quenching. The binding constants Ka at 300, 310, 320 K are separately 6.93×1012,7.40×1012,6.82×1012 L/mol/s respectively. Number of binding sites n is 1; the binding distance R is 3.70 nm, and that suggests non-radiation energy transfer from BSA to pheophorbide. The thermodynamic parameters of the binding reaction are H=36.7 kJ/mol, S=213 J/mol/K, and G negative value, and indicates that hydrophobic force plays a predominant role in the process, and it is a spontaneous interaction. Synchronous fluorescence spectra show that pheophorbide mainly interacts with tryptophan residue of BSA and leads to the promotion of hydrophobic force. Pheophorbide can bind to serum protein and be transported in vivo, makes no destruction to molecular structure of serum protein, but causes its conformational alteration.

Improvements in a Chemically Defined Medium for the Growth of Mouse Cells (Strain LS) in Suspension

1970 ◽  
Vol 7 (3) ◽  
pp. 661-670
Author(s):  
J. R. BIRCH ◽  
S. J. PIRT
Keyword(s):  
Amino Acids ◽  
Population Density ◽  
Cell Population ◽  
Defined Medium ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Free Medium ◽  
Average Maximum ◽  
Maximum Cell ◽  
Medium Change

In the medium used in previous work, both growth rate and maximum cell population density were reduced in protein-free medium. This was due to iron limitation. In a medium supplemented with iron, the average minimum population doubling time was reduced from 78 to 37 h and the average maximum cell population density increased from 1.6x106 to 3.3x106/ml. In the improved medium described, sodium pyruvate, α-ketoglutaric acid, methylcellulose and polyvinylpyrrolidone are no longer required although methylcellulose prevents an initial fall in population which occurs in polymer-free medium. The present study indicates that amino acids now limit maximum cell population density. By increasing the concentration of the relevant amino acids we have obtained maximum cell populations in excess of 5.106 cells/ml without medium change. Finally we describe a method for measuring the growth of the LS cell in the defined medium by an opacity method.

scholarly journals In silico and multi-spectroscopic analyses on the interaction of 5-amino-8-hydroxyquinoline and bovine serum albumin as a potential anticancer agent

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Waralee Ruankham ◽  
Kamonrat Phopin ◽  
Ratchanok Pingaew ◽  
Supaluk Prachayasittikul ◽  
Virapong Prachayasittikul ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Serum Protein ◽  
In Silico ◽  
Bovine Serum ◽  
Electrostatic Interactions ◽  
Water Solubility ◽  
Free Fraction ◽  
Amino Derivative

Abstract5-Amino-8-hydroxyquinoline (5A8HQ), an amino derivative of 8-hydroxyquinoline, has become a potential anticancer candidate because of its promising proteasome inhibitory activity to overcome and yet synergize bortezomib for fighting cancers. Therefore, in this study, its physicochemical properties and interaction activities with serum protein have extensively been elucidated by both in vitro and in silico approaches to fulfill the pharmacokinetic and pharmacodynamic gaps. 5A8HQ exhibited the drug-likeness properties, where oral administration seems to be a route of choice owing to its high-water solubility and intestinal absorptivity. Multi-spectroscopic investigations suggested that 5A8HQ tended to associate with bovine serum albumin (BSA), a representative of serum protein, via the ground-state complexation. It apparently bound in a protein cleft between subdomains IIA and IIIA of BSA as suggested by the molecular docking and molecular dynamics simulations. The binding was mainly driven by hydrogen bonding and electrostatic interactions with a moderate binding constant at 104 M−1, conforming with the predicted free fraction in serum at 0.484. Therefore, 5A8HQ seems to display a good bioavailability in plasma to reach target sites and exerts its potent pharmacological activity. Likewise, serum albumin is a good candidate to be reservoir and transporter of 5A8HQ in the circulatory system.

Improvement of an artificial test substrate technique for evaluation of em immunocytochemical labeling

1987 ◽  
Vol 45 ◽  
pp. 762-763
Author(s):  
G. D. Gagne ◽  
M. F. Miller
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Artificial Substrate ◽  
Chemical Fixation ◽  
As If

We recently described an artificial substrate system which could be used to optimize labeling parameters in EM immunocytochemistry (ICC). The system utilizes blocks of glutaraldehyde polymerized bovine serum albumin (BSA) into which an antigen is incorporated by a soaking procedure. The resulting antigen impregnated blocks can then be fixed and embedded as if they are pieces of tissue and the effects of fixation, embedding and other parameters on the ability of incorporated antigen to be immunocyto-chemically labeled can then be assessed. In developing this system further, we discovered that the BSA substrate can also be dried and then sectioned for immunolabeling with or without prior chemical fixation and without exposing the antigen to embedding reagents. The effects of fixation and embedding protocols can thus be evaluated separately.

Stability of Prostacyclin in Human Plasma and Whole Blood: Studies on the Protective Effect of Albumin

1981 ◽  
Vol 46 (03) ◽  
pp. 645-647 ◽  
Author(s):  
M A Orchard ◽  
C Robinson
Keyword(s):  
Platelet Aggregation ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Protective Effect ◽  
Whole Blood ◽  
Bovine Serum ◽  
Fatty Acid Content ◽  
Krebs Solution ◽  
Antiaggregatory Activity ◽  
Free Plasma

SummaryThe biological half-life of prostacyclin in Krebs solution, human cell-free plasma or whole blood was measured by bracket assay on ADP-induced platelet aggregation. At 37°C, pH 7.4, plasma and blood reduced the rate of loss of antiaggregatory activity compared with Krebs solution. The protective effect of plasma was greater than that of whole blood. This effect could be partially mimicked by the addition of human or bovine serum albumin to the Krebs solution. The stabilisation afforded by human serum albumin was dependent on the fatty acid content of the albumin, although this was less important for bovine serum albumin.

RADIOIMMUNOASSAY OF OESTRONE AND OESTRADIOL IN THE PERIPHERAL PLASMA OF THE DOMESTIC FOWL IN VARIOUS PHYSIOLOGICAL STATES AND OF HYPOPHYSECTOMIZED AND OVARIECTOMIZED FOWLS

1974 ◽  
Vol 75 (1) ◽  
pp. 133-140 ◽  
Author(s):  
B. E. Senior
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Diethyl Ether ◽  
Bovine Serum ◽  
Domestic Fowl ◽  
Laying Hens ◽  
Peripheral Plasma ◽  
The Mean ◽  
Precision And Accuracy ◽  
Chicken Ovary

ABSTRACT A radioimmunoassay was developed to measure the levels of oestrone and oestradiol in 0.5–1.0 ml of domestic fowl peripheral plasma. The oestrogens were extracted with diethyl ether, chromatographed on columns of Sephadex LH-20 and assayed with an antiserum prepared against oestradiol-17β-succinyl-bovine serum albumin using a 17 h incubation at 4°C. The specificity, sensitivity, precision and accuracy of the assays were satisfactory. Oestrogen concentrations were determined in the plasma of birds in various reproductive states. In laying hens the ranges of oestrone and oestradiol were 12–190 pg/ml and 29–327 pg/ml respectively. Levels in immature birds, in adult cockerels and in an ovariectomized hen were barely detectable. The mean concentrations of oestrone and oestradiol in the plasma of four non-laying hens (55 pg/ml and 72 pg/ml respectively) and one partially ovariectomized hen (71 pg/ml and 134 pg/ml respectively) were well within the range for laying hens. It is evident that the large, yolk-filled follicles are not the only source of oestrogens in the chicken ovary.

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