+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Adjustment of the osmolality of Percoll for the isopycnic separation of cells and cell organelles

, : Adjustment of the osmolality of Percoll for the isopycnic separation of cells and cell organelles. Analytical Biochemistry 141(2): 322-328

The addition of 1 part of 1.5 M NaCl or 2.5 M sucrose (10 X concentrate) to 9 parts of Percoll produces a stock solution that is hypertonic (350-360 mOsm/kg H2O). Because the osmolality is a critical variable in the isopycnic separation of cells and cell organelles, the factors accountable for this hypertonicity were investigated. Percoll, a colloidal suspension of silica particles coated with polyvinylpyrrolidone, can be described as a medium composed of two distinct compartments, an aqueous phase and a solid phase. According to this model, solutes (e.g., NaCl, sucrose) should have access to the aqueous phase and add to the intrinsic osmolality of Percoll, but should be excluded from the solid phase. In order to verify this hypothesis, mathematical equations were derived and tested. It was found that the ratio of the aqueous volume to the total volume of Percoll (R value) was dependent on the ionic strength of the stock solution. With this parameter, the osmolality of Percoll stock solutions could be predicted (+/-2%) and, consequently, one could calculate the proper dilution to be used with saline, culture medium, or sucrose concentrates to obtain a truly isotonic Percoll stock solution (congruent to 290 mOsm/kg H2O). The relative importance of an accurate control of the osmolality in the preparation of density gradients made up of Percoll is also discussed.


PMID: 6093627

Other references

Ayeni, J.; Ayanda, J., 1982: Studies of the husbandry practices and social acceptance of guineafowl in Nigeria. Bulletin of animal health and production in Africa = Bulletin des sante et production animales en Afrique 30(2): 139-148

Bityukova Liidia; Shogenova Alla, 1996: Comparative analysis of geochemical processes in the early Paleozoic sedimentary basin of Estonia. International Geological Congress, Abstracts--Congres Geologique Internationale, Resumes 30, Vol

Lillie, R.D., 1944: Factors influencing the Romanovsky staining of blood films and the role of methylene violet. The constitution of the azure component of Wright's stain is difficult to control when the usual process of steaming one hour with NaHCO3is used. Alkali polychroming methods are difficult to control but acid polychroming with 200 mg. K3Cr2O7...

John, L.Jenny.; Ahmed, S.; Anjum, F.; Kebab, M.; Mohammed, N.; Darwich, H.; Ibraheem, N.; Arifulla, M.; Sreedharan, J., 2014: Prevalence of Allergies among University Students: A Study from Ajman, United Arab Emirates. Aim. Urbanization and globalization in the Middle East have resulted in drastic environmental changes and increased allergens present in the environment. This study aimed to assess the prevalence of allergies among undergraduate students from a un...

Croxatto H.B.; Ortiz M.E.; Forcelledo M.L.; Fuentealba B., 1982: Ovum transport and its regulation. Archivos de Biologia y Medicina Experimentales 15(3-4): 309-314

Moodley, J.; Norman, R.J.; Reddi, K., 1984: Central venous concentrations of immunoreactive prostaglandins E, F, and 6-keto-prostaglandin F1 in eclampsia. Concentrations of prostaglandins E, F, and 6-keto-prostaglandin F1 alpha were estimated in central venous blood and amniotic fluid in 21 women with eclampsia and 16 healthy pregnant controls. Central venous blood concentrations of 6-keto-prostagla...

Adolf G.; Bischoff H M.; Mueller M., 1989: Effect of seed mixture and sowing method on yielding potential of swards established on a bottom land site along the middle course of the river elbe. Site-adapted grass mixtures grown along the middle course of the River Elbe gave dry-matter yields of more than 10 t/ha in their first year of cutting. In the second year the sowing effect vanished as the yielding potential of the high-yielding va...

Filippi, L.; Biancone, L.; Petruzziello, C.; Schillaci, O., 2006: Tc-99m HMPAO-labeled leukocyte scintigraphy with hybrid SPECT/CT detects perianal fistulas in Crohn disease. Clinical Nuclear Medicine 31(9): 541-542

Cerezo-Guisado, M.Isabel.; Garcia-Marin, L.Jesus.; Lorenzo, M.Jesus.; Bragado, M.Julia., 2005: Lovastatin inhibits the growth and survival pathway of phosphoinositide 3-kinase/protein kinase B in immortalized rat brain neuroblasts. We previously showed that lovastatin, an HMG-CoA reductase inhibitor, suppresses cell growth by inducing apoptosis in rat brain neuroblasts. Our aim was to study intracellular signalling induced by lovastatin in neuroblasts. Lovastatin significant...

Anonymous, 1952: Preparation of cortisone. Rassegna Giuliana di Medicina 8(10-12): 221-223