Fusion-tag affinity chromatography is a key technique in recombinant protein purification. Current methods for protein recovery from mammalian cells are hampered by the need for feed stream clarification. We have developed a method for direct capture using immobilized metal affinity chromatography (IMAC) of hexahistidine (His6) tagged proteins from unclarified mammalian cell feed streams. The p...

We have made Chinese hamster ovary (CHO) cell lines that contain up to 2000 copies of the coding region of the mouse c-myc gene fused to the promoter of the Drosophila gene (hsp70) encoding a Mr 70,000 heat shock protein. Incubation of these cells at 43 degrees C results in an estimated 100-fold induction of c-myc mRNA. Translation of this mRNA occurs when the cells are returned to 37 degrees C...

Translationally Controlled Tumour Protein (TCTP) associates with microtubules (MT), however, the details of this association are unknown. Here we analyze the relationship of TCTP with MTs and centrosomes in Xenopus laevis and mammalian cells using immunofluorescence, tagged TCTP expression and immunoelectron microscopy. We show that TCTP associates both with MTs and centrosomes at spindle poles...

A novel culture system for mammalian cells was used to investigate division orientations in populations of Chinese hamster ovary cells and the influence of gravity on the positioning of division axes. The cells were tethered to adhesive sites, smaller in diameter than a newborn cell, distributed over a nonadhesive substrate positioned vertically. The cells grew and divided while attached to the...

Endocannabinoid synthesizing 2-AG and cannabinoid receptors have been reported in some organ of reproductive system oocyte ovary, but little is known about the expression subcellular localization CB1 DAGL-α interstitial gland tissue mouse ovary. The aim this study was to explain differential distribution endocannabinoid glandular cell DAGL-α-immunoreactivities were detected within cells, which ...

BACKGROUND The majority of oocytes in the mammalian ovary are dormant oocytes that are enclosed in primordial follicles by several somatic cells, which we refer to as primordial follicle granulosa cells (pfGCs). Very little is known, however, about how the pfGCs control the activation of primordial follicles and the developmental fates of dormant oocytes. RESULTS By targeting molecules in pfG...

Toxoplasmosis is a worldwide infection which is commonly asymptomatic but can cause serious medical problems in immunocompromised individuals and fetus. The infection also causes considerable economic loss because of abortion in livestock, mostly in sheep and goats. DNA vaccination may be a powerful approach against intracellular parasites such as Toxoplasma gondii. The goal of this study was t...

Background This novel approach describes a rapid and simple method for identification of necrotic vs. viable cells within a mammalian blastocyst. MaterialsAndMethods Hatched bovine blastocysts produced in vitro were first incubated for 30 min in pre-equilibrated culture medium containing propidium iodide (PI; 300μg/ml) and bisbenzimide (Hoechst: H33342; 5μg/ml) fluorescent dyes. Embryos were th...

-l. The localization of radiolabeled human chorionic gonadotrophin (~2SI-HCG) was studied in a number of commonly used mammalian animal models. 2. The ovary proved to be the target organ for ~25I-HCG in all animals studied. 3. The optimum time for the localization of HCG in the ovary varied in all the mammals extending from 1.5 to 4 hr. 4. HCG is apparently used in the target cell for only a br...

objective: the possible toxicity of drugs in pregnancy should be tested before their use in pregnant patients. in the present study, we aimed to evaluate the safety of phytovagex, a pessary formulation of nigella sativa (n. sativa), which is already in clinical use for vaginal fungal infection. materials and methods: the pregnant rats were treated intravaginal with physiological saline (vehicle...