Percoll®
Referência sc-500790A
Tamanho : 100ml
Marca : Santa Cruz Biotechnology
Percoll®
ACCÈS RAPIDE AUX LIENS
Percoll® est utilisé dans les applications de centrifugation de densité pour la séparation des cellules, des particules subcellulaires et des virus de grande taille (jusqu'à ~ 70S) dans des conditions douces qui préservent la viabilité et l'intégrité morphologique. Percoll® est constitué de particules de silice (15 à 30 nm de diamètre) enrobées de polyvinylpyrrolidone (PVP) non dialysable. Le PVP libre n'est présent qu'à hauteur de 1 à 2 %. Percoll® est non toxique, presque chimiquement inerte et n'adhère pas aux membranes. Percoll® est ajustable à la force ionique et au pH physiologiques, et les gradients peuvent être réalisés ou générés spontanément par centrifugation à des vitesses modérées dans un rotor à tête inclinée. Les gradients sont iso-osmotiques et couvrent une gamme de densités allant jusqu'à 1,3 g/ml. Percoll® a également été utilisé dans la préparation du sperme pour étudier les besoins énergétiques des embryons bovins.
Percoll® Références:
- Fractionation of cells and subcellular particles with Percoll. | Pertoft, H. 2000. J Biochem Biophys Methods. 44: 1-30. PMID: 10889273
- A simple method for selection of cumulus-oocyte complexes from bovine ovaries by sedimentation with percoll. | Yotsushima, K., et al. 2007. J Reprod Dev. 53: 971-6. PMID: 17519521
- Isolation of lysosomes from tissues and cells by differential and density gradient centrifugation. | Graham, JM. 2001. Curr Protoc Cell Biol. Chapter 3: Unit 3.6. PMID: 18228358
- Optimal Percoll concentration facilitates flow cytometric analysis for annexin V/propidium iodine-stained ischemic brain tissues. | Juan, WS., et al. 2012. Cytometry A. 81: 400-8. PMID: 22311734
- Purification Toxoplasma gondii Tissue Cysts Using Percoll Gradients. | Watts, EA., et al. 2017. Curr Protoc Microbiol. 45: 20C.2.1-20C.2.19. PMID: 28510363
- Using Percoll Gradient Fractionation to Study the Endocytic Trafficking of the EGFR. | Gosney, JA. and Ceresa, BP. 2017. Methods Mol Biol. 1652: 145-158. PMID: 28791641
- Gradient sperm selection for reproductive techniques in cattle: Is Isolate a suitable replacement for Percoll? | Sepúlveda, B., et al. 2018. Andrologia. 50: PMID: 29164653
- Isolation and Purification of Satellite Cells from Young Rats by Percoll Density Gradient Centrifugation. | Matsuyoshi, Y., et al. 2019. Methods Mol Biol. 1889: 81-93. PMID: 30367410
- Impact of Percoll purification on isolation of primary human hepatocytes. | Horner, R., et al. 2019. Sci Rep. 9: 6542. PMID: 31024069
- Porcine IVF embryo development and estrogen receptors are influenced by the concentration of percoll gradients during sperm selection. | Ohlweiler, LU., et al. 2020. Mol Reprod Dev. 87: 135-141. PMID: 31691431
- Purification and production of Plasmodium falciparum zygotes from in vitro culture using magnetic column and Percoll density gradient. | Zhou, Y., et al. 2020. Malar J. 19: 192. PMID: 32450861
- Purification of Functional Platelet Mitochondria Using a Discontinuous Percoll Gradient. | Léger, JL., et al. 2021. Methods Mol Biol. 2276: 57-66. PMID: 34060032
- Purification of Leaf Mitochondria from Arabidopsis thaliana Using Percoll Density Gradients. | Tran, HC. and Van Aken, O. 2022. Methods Mol Biol. 2363: 1-12. PMID: 34545481
- Isolation of Highly Purified, Intact, and Functional Mitochondria from Potato Tubers Using a Two-in-One Percoll Density Gradient. | Møller, IM. and Rasmusson, AG. 2022. Methods Mol Biol. 2363: 39-50. PMID: 34545484
- Continuous Percoll Gradient Centrifugation of Erythrocytes-Explanation of Cellular Bands and Compromised Age Separation. | Maurer, F., et al. 2022. Cells. 11: PMID: 35455975
⋅ Percoll® can be buffered within the pH range 5.5 to 10.0 without any changes in properties. If the pH is dropped below 5.5, gelling may occur. Gelling can also be caused by the presence of divalent cations, an effect which is exacerbated by elevated temperatures.
⋅ Undiluted Percoll® can be resterilized by autoclaving for 30 min at 120°C.
Preparation of gradient material
Percoll® PLUS/Percoll® are best used in balanced salt solutions, physiological saline, or 0.25 M sucrose. Cells can be separated on gradients in balanced salt solutions. Subcellular particles, however, tend to aggregate in the presence of salts and it is recommended that the separation of such particles be carried out in Percoll®diluted with sucrose (0.25 M final concentration).
The low osmolality of Percoll® permits this parameter to be controlled by the user without significant interference from the density medium itself. The addition of 9 parts (v/v) of Percoll® to 1 part (v/v) of either 1.5 M NaCl, 10x concentrated cell culture medium, or 2.5 M sucrose will result in a solution adjusted to about 340 mOsm/kg H2O. Solutions of different osmotic pressure can be produced by adjusting the relative volumes of Percoll® and salt or sucrose solution. The final adjustment to the required osmolality can be carried out by the addition of salts or distilled water. When precise osmotic pressures are required, it is recommended that the osmolality of the solutions be measured in an osmometer.
Centrifugation with Percoll®
Percoll® will form self-generated gradients by
centrifugation at approximately 10,000 gav (in 0.15 M saline) or 25,000 gav (in 0.25 M sucrose) in fixed-angle rotor heads after 15 minutes. Cells or subcellular particles can be mixed with Percoll® prior to centrifugation and will band isopycnically, as the gradient is formed in situ. Although Percoll® is best used in angle-head rotors, banding of cells on pre-formed (continuous or discontinuous) gradients may be carried out at 400 gav for 20 to 30 minutes in swing-out rotors.
Removal of Percoll®after centrifugation
To remove the gradient medium from the biological material, perform one of the procedures outlined below.
• Cells can be recovered free from particles of Percoll® by dilution with physiological saline and centrifugation to collect the cells.
• Subcellular particles can be separated from Percoll® by the procedure described above. The size of the particles will determine the centrifugal force required to separate the particles from Percoll®.
• Gel filtration or ion exchange chromatography can also be used to separate biological material from Percoll®.
Aggregates of silica particles
It is an inherent tendency of all silica colloids to form aggregates on prolonged storage. These aggregates may be observed in some batches of Percoll®, either as a slight sediment at the bottom of the tube or as a faint white band with a density of 1.04 to 1.05 g/ml. This band may form during gradient formation in the centrifuge or during low speed centrifugation of a performed gradient. The aggregated silica does not interfere with the separation of biological particles and almost all cells and organelles have buoyant densities of greater than 1.05 g/ml in PPercoll® PLUS For the majority of cell, virus, and organelle separations, any silica aggregates banded from the gradient material (see above) may be ignored.
For specific experiments, it may be desirable to remove aggregates; this may be achieved by filtration of Percoll® through a depth filter prior to centrifugation.
Aggregation is not a problem in Percoll® PLUS) due to the silane coating.
Formulation :
Colloidal solution of silica coated with polyvinylpyrrolidone (PVP)
Apparence :
Suspension
État Physique :
Liquid
pH :
8.5 - 9.5
Stockage :
Store at room temperature
Densité :
max: 1.135 g/mL
Télécharger la Fiche de Sécurité/ SDS (MSDS)
Région
EXEMPLE de Certificat d'Analyse (CoA)
Percoll® SAMPLE Certificate of AnalysisCERTIFICAT D'ANALYSE
Numéro de catalogue
Numéro de Lot
SAMPLE Certificate of Analysis (COA)
Test | Specification | Results |
---|---|---|
Conductivity | ≤100 mS/m | 64 mS/m |
Density | 1.125 - 1.135 g/mL | 1.130 g/mL |
pH | 8.5 - 9.5 | 9.0 |
Viscosity | ≤15 cP | 12 cP |
Osmolality | ≤25 mOsmol/kg | 15 mOsmol/kg |
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Informations pour la commande
Nom du produit | Ref. Catalogue | COND. | Prix HT | QTÉ | Favoris | |
Percoll®, 25 ml | sc-500790 | 25 ml | .00 | |||
Percoll®, 100 ml | sc-500790A | 100 ml | .00 |