The intracellular localization of the activity and synthesis of three isozymes of NAD(P)(+)-glutamate dehydrogenase from the unicellular green alga Chlamydomonas reinhardtii cw-92 has been established. Isozyme activities have been located within mitochondria by using differential centrifugation techniques and discontinuous Percoll gradient separations. Experiments with protein synthesis inhibit...

Surface immunoglobulin D (IgD)-positive lymphocytes precoated with monoclonal anti-delta antibody were selectively removed from blood mononuclear cell preparations by "panning" and by fluorescence-activated cell sorter. The depletion of sIgD+ cells did not significantly affect plasma cell responses to pokeweed mitogen PWM). PWM-responsive B cells lacking sIgD and mouse erythrocyte receptors pre...

Coat protein (CP) of Hibiscus chlorotic ringspot virus (HCRSV) had been found to be responsible for the spread of infection. Kenaf (Hibiscus cannabinus) was used as a model organism in this study. As CP is most likely localized in the peroxisomes, there is a need to produce a protocol adapted solely for the isolation of peroxisomes from the leaf of Kenaf as peroxisomal characteristics differ gr...

Background Sexing sperm is an important biotechnical that enables improve animal production, with achievement of maximum animal breeding. However, all sex semen methodologies can promote irreparable damages to sperm cell, which compromise their fertility [1]. Among the main causes of injures to the sperm are the reactive oxygen species, which have their production intensified during semen manip...

Rat peritoneal mast cells obtained by Percoll gradient were incubated with different concentrations of naturally occurring aliphatic polyamines, spermine and spermidine, at 0.1-10 mM and the amount of histamine release into the supernatant solutions was measured. The addition of each polyamine to the suspensions of the mast cells caused a histamine release in a dose-dependent manner. The histam...

Intact leucoplasts from the endosperm of developing castor oil seed were isolated by Percoll density gradient centrifugation. The precursor to the small subunit of ribulose 1,5-bisphosphate carboxylase from pea was synthesized in vitro from hybrid-selected mRNA. Leucoplasts imported this precursor by an ATP-requiring mechanism similar to that described in chloroplasts (AR Grossman et al. 1980 N...

A cellular compartment from brown adipose tissue (BAT) of newborn rats was isolated by Percoll-density-gradient centrifugation and was shown to proliferate and to undergo adipose conversion in vitro in primary culture. The features of the effector requirement for adipose conversion as well as the differentiated morphological and biochemical phenotype are almost identical with that of a compartm...

The mitochondrion of the parasitic protozoon Trypanosoma brucei shows a number of unique features, many of which represent highly interesting research topics. Studies of these subjects require the purification of mitochondrial fractions. Here, we describe and discuss the two most commonly used methods to isolate mitochondria from insect stage T. brucei. In the first protocol, the cells are lyse...