A simple method for human peripheral blood monocyte isolation

We describe a simple method using percoll gradient for isolation of highly enriched human monocytes. High numbers of fully functional cells are obtained from whole blood or buffy coat cells. The use of simple laboratory equipment and a relatively cheap reagent makes the described method a convenient approach to obtaining human monocytes.     

A fast method for high-quality genomic DNA extraction from whole human blood

A simple and fast protocol is described for the purification of genomic DNA from 0.3 ml of whole human blood. The recovery of DNA is quantitative and reproducible; the quality is such that it can be used for all relevant molecular biology techniques.     

A new simple method of preparing actin from chicken gizzard

A simple method for preparing actin from chicken gizzard was described. This method takes advantage of a property of gizzard tropomyosin, that is, that it does not form Mg paracrystals readily.     

An improved method for preparing whole specimens from bovine preimplantation embryos: A technique note

A method was devised to prevent loss of whole embryos during fixation. Specimens were prepared in a chamber saturated with fixative vapors consisting of 3 : 1 (v/v) 96%. ethanol/glacial acetic acid. Good quality specimens were obtained after fixation for at least 24 but not more than 72 h. After staining, specimens could be preserved for 3 to 4 d by storage in the fixation chamber, in 45% aqueous acetic acid vapor. Using the method suggested in this paper prevents loss of early embryos during fixation and allows storage of specimens for longer than usual time while maintaining the quality of the specimen.     

A new synthetic method for preparing indole derivatives from 2-keto glycosides

In this article, a new reaction of the addition of two molecules of aniline to 2-keto glycosides (glycoside 2-uloses, 2-ulosides) is reported. A possible pathway for the reaction is presented. This reaction provides a novel one-pot method for the synthesis of indole derivatives from sugars.     

Monocyte separation from whole blood : A method for experimental animals

A large supply of monocytes can be obtained from whole blood of rats. The principle is to stimulate a monocytosis, separate the lymphocyte/monocyte series on a Ficoll/sodium metrizoate gradient and eliminate lymphocytes with anti-lymphocyte serum. A simple method of labelling monocytes with tritiated thymidine is described. Some possible situations for studying monocytes in syngeneic animals in experimental situations are mentioned.     

A method for whole protein isolation from human cranial bone

The presence of the dense hydroxyapatite matrix within human bone limits the applicability of conventional protocols for protein extraction. This has hindered the complete and accurate characterization of the human bone proteome thus far, leaving many bone-related disorders poorly understood. We sought to refine an existing method of protein extraction from mouse bone to extract whole proteins of varying molecular weights from human cranial bone. Whole protein was extracted from human cranial suture by mechanically processing samples using a method that limits protein degradation by minimizing heat introduction to proteins. The presence of whole protein was confirmed by western blotting. Mass spectrometry was used to sequence peptides and identify isolated proteins. The data have been deposited to the ProteomeXchange with identifier PXD003215. Extracted proteins were characterized as both intra- and extracellular and had molecular weights ranging from 9.4 to 629 kDa. High correlation scores among suture protein spectral counts support the reproducibility of the method. Ontology analytics revealed proteins of myriad functions including mediators of metabolic processes and cell organelles. These results demonstrate a reproducible method for isolation of whole protein from human cranial bone, representing a large range of molecular weights, origins and functions.     

人血基因组PCR扩增模板的制血及其保存

建立一种简单的人血基因组PCR反应扩增模板制备及其保存方法,用于α-globin基因HS40位点扩增。     

A new method of preparing hemin conjugates with rabbit IgG

Stable derivatives of hemin activated by means of the Woodward reagent K were obtained, and the reaction conditions for their covalent attachment to the IgG molecule optimized. Advantages of the new method for conjugation are discussed.     

A new method of preparing gold probes for multiple-labeling cytochemistry

A new method is described for preparing colloidal gold particles in any size between approximately 3 and 17 nm for electron microscopy. The gold particles are homogeneous in size (homodisperse). When bound to various proteins (e.g. IgG and protein A), the complexes were stable for long periods and suitable for affinity cytochemistry. We demonstrated the usefulness of the new gold probes bound to protein A for multiple labeling in a current immunocytochemical study on receptor mediated transport of IgA in human duodenal crypt cells. Other gold-protein complexes were useful for studying macromolecular arrangements at high resolution.     

A new capillary viscometer for whole blood viscosimetry

A new capillary system was developed, incorporating infrared sensors, which allowed the determination of whole blood viscosity over a wide range of shear stresses. Flow conditions were defined by the geometry of the capillary and the sample pressure head. Whole blood was considered to be a power law fluid and a modified Mooney's formula was used for the calculation of the related invariants. The new viscometer proved to be very simple in use, requiring one run, had a short measuring time and utilised a small test sample volume. However it can be used for whole blood viscosity measurements only at medium and high shear stresses.     

An improved method for preparing G-banded chromosomes from mouse peripheral blood

We describe here several improvements in the method we originally developed to prepare mitotic chromosomes from peripheral blood of laboratory mice. In addition, we have tried several methods to improve metaphase yield from lymphocytes of the inbred strain DBA/2J, which respond poorly to phytohemagglutinin. The yield of mitoses from DBA/2J cells cannot be improved by enhancing T-cell response using interleukin-2 or by using a different T-cell mitogen, concanavalin A. Metaphase yield from peripheral blood cells of DBA/2J mice can be improved significantly by adding lipopolysaccharide to cultures, probably stimulating B-cell as well as T-cell proliferation.     

Thixotropic properties of whole blood from healthy human subjects

The steady state non-Newtonian viscosity of whole human blood has been widely studied as a function of the shear rate; and used to characterize the blood in various pathological disorders. In our previous studies, we demonstrated that blood is a thixotropic fluid. Its time-dependency and shear rate dependency of rheological behavior can be represented by an equation developed by Huang. Parameters of the equation can be used for the characterization of an individual's blood. They provide information, such as the kinetic rate constant of breakdown of RBC rouleaux to individual erythrocytes and the relative amount of rouleau formation in the dynamic equilibrium between rouleaux and individual erythrocytes. In this communication, the thixotropic parameters from blood samples of fifteen apparently healthy human subjects were investigated. When compared to the use of apparent viscosity values for the correlation with a pathological disorder, thixotropic parameters are preferable. The mean values of thixotropic parameters obtained from apparently healthy human subjects provide a base for comparison with the same parameters as obtained from blood samples of patients with certain pathological disorders involving the circulation.