HiMedia Laboratories - culture media - CADERSKY-ENVITEK Ltd.,Czech Republic, Europe

CADERSKY-ENVITEK Ltd.

Brno, Czech Republic Europe



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HiSep™

HiSep™ LSM 1077 Sterile filtered solution containing polysucrose and sodium diatrizoate, having a density 1.0770 ±0.0010. It facilitates the recovery of large numbers of viable human mononuclear cells Centrifugation causes mononuclear cells to form a distinct layer at the plasma- HiSep™ LSM 1077 interface. HiSep™ LSM 1077 is equivalent to Ficoll Hypaque™ and Histopaque™-1077.	LS001-100ML LS001-6x100ML LS001-500ML LS001-6x500ML LS001-12x500ML	100ml 6X100ml 500ml 6x500ml 12x500ml
HiSep™ LSM 1073 Sterile filtered solution similar to HiSep LSM 1077, adjusted to a density of 1.073 g/ml. It facilitates isolation of lower density human mononuclear cells (eg- Mesenchymal stromal cells or monocytes).	LS002-100ML LS002-6X100ML LS002-500ML LS002-6X500ML LS002-12X500ML	100ml 6X100ml 500ml 6X500ml 12X500ml
HiSep™ LSM 1084 Sterile filtered solution containing polysucrose and sodium diatrizoate, having a density of 1.084 ± 0.001 g/ml. It facilitates the recovery of large numbers of viable mononuclear cells from chicken, rats, mice and other mammals. Centrifugation causes mononuclear cells to form a distinct layer at the plasma-HiSep™ LSM 1084 interface. HiSep™ LSM 1084 is equivalent to Ficoll-Paque PREMIUM 1.084.	LS003-100ML LS003-6X100ML LS003-500ML LS003-6X500ML LS003-12X500ML	100ml 6X100ml 500ml 6X500ml 12X500ml
GranuloSep™ GSM 1119 Sterile filtered solution similar to HiSep™ LSM 1077, adjusted to a density of 1.119 g/ml. When combined with HiSep™ LSM 1077, it permits the separation of mononuclear cells and granulocytes. Solution is for in vitro diagnostic use. GranuloSep™ GSM 1119 is equivalent to Histopaque-1119.	LS004-100ML LS004-6X100ML LS004-500ML LS004-6x500ML	100ml 6X100ml 500ml 6x500ml
Ficoll 400-20% A 0.2 micron filtered, non-ionic synthetic polymer of sucrose. It is used in electrophoresis and as a hapten carrier. Most commonly used to prepare density gradients	LS005-25ML LS005-50ML LS005-500ML	25ml 50ml 500ml

LS001 Lymphocyte Separation Media 1077 The first step in studying lymphocytes (present in blood, peritoneal exudates or lymphoid organs mixed with other cells) is to isolate them so that their behavior can be analyzed in vitro. HiSep™ LSM 1077 is based on the adapted method of isolating lymphocytes using centrifugation techniques by Boyum in which diluted defibrinated blood is layered on a solution of sodium diatrizoate and polysucrose (adjusted to a density of 1.0770 ± 0.0010 g/ml) and centrifuged at low speeds for 30 minutes. This medium offers a quick and reliable method for the simple isolation of human mononuclear cells and lymphocytes from defibrinated EDTA or heparin-treated human blood. It is certified for in vitro Diagnostic (IVD) use. The method is applicable for studying cell-mediated lympholysis and for human lymphocyte antigen (HLA) typing. It may also be employed as the initial step prior to enumeration of T-, B- and “null” lymphocytes.

Replace other Expensive Lymphocyte Separation Medium with Cost effective Premium Quality HiSep™ LSM 1077

Figure 1. Differential migration following centrifugation results in the formation of several cell layers. Mononuclear cells (lymphocytes and monocytes) and platelets are contained in the banded plasma-LSM interphase due to their density, and the pellet that is formed contains mostly erythrocytes and granulocytes, which have migrated through the gradient to the bottom of the tube. Lymphocytes are recovered by aspirating the plasma layer and then removing the cells. Excess platelets, HiSep™ LSM 1077, and plasma can then be removed by cell washing with isotonic phosphate buffered saline.

Easy to perform, minimal steps and handy protocol

Aseptically transfer 2.5 ml of HiSep™ LSM 1077 to a 15 ml clean centrifuge tube and overlay with 7.5 ml diluted blood.
Centrifuge at 400 x g with the brake off, at room temperature for 15-30 minutes. Centrifugation should sediment erythrocytes and polynuclear leukocytes and band mononuclear lymphocytes above HiSep™ LSM 1077 as shown in Fig.1.
Aspirate most of the plasma and platelet containing supernatant above the interphase band (granulocytes and erythrocytes will be in the red pellet). Using a clean glass pasteur pipette carefully aspirate the lymphocyte layer along with half of the HiSep™ LSM 1077 layer.
Two washes with isotonic phosphate buffered saline separates the cell from separation medium.
Count the cells and also determine the number of viable cells by trypan blue exclusion staining.