Description
General description
ACCUSPIN System-Histopaque-1077 products are intended for use in the isolation of lymphocytes and other mononuclear cells. The separation medium, Histopaque-1077, is a sterile-filtered, endotoxin-tested solution of polysucrose and sodium diatrizoate, adjusted to a density of 1.077 g/mL. ACCUSPIN System-HISTOPAQUE-1077 employs γ-irradiated 50 mL polypropylene centrifuge tubes specially designed with two chambers separated by a porous high-density polyethylene barrier ("frit"). Each tube is aseptically filled with 15 mL of Histopaque 1077. Each tube will separate 15-30 mL of anticoagulated blood.
Application
ACCUSPIN System-HISTOPAQUE-1077 employs centrifuge tubes specially designed with two chambers separated by a porous high-density polyethylene barrier ("frit"). The lower chamber contains HISTOPAQUE-1077 which allows the addition of anticoagulated whole blood without risk of mixing with the separation medium. On centrifugation, the whole blood descends through the frit to contact with the HISTOPAQUE-1077 below the frit, giving a clear separation of the blood components. The erythrocytes aggregate and the granulocytes become slightly hypertonic, increasing their sedimentation rate, resulting in pelleting at the bottom of the ACCUSPIN tube. Lymphocytes and other mononuclear cells, i.e., monocytes, remain at the plasma-HISTOPAQUE-1077 interface.
ACCUSPIN System-Histopaque-1077 is used for the separation of lymphocytes and other mononuclear cells from whole blood and bone marrow using density gradient separation media. Histopaque-1077 has been used for:
human lymphocyte antigen (HLA) typing
initial isolation step prior to the enumeration of T, B, and ‘null’ lymphocytes.
preparation of pure lymphocyte suspensions for cell culture and cytotoxicity assays.
isolation and purification of peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMCs)citation
isolation of human lymphocytescitation
isolation of breast cancer circulating tumor cells (CTCs) from the whole bloodcitation
Biochem/physiol Actions
The lower chamber contains HISTOPAQUE-1077 which allows the addition of anticoagulated whole blood without risk of mixing with the separation medium. On centrifugation, the whole blood descends through the frit to contact with the HISTOPAQUE-1077 below the frit, giving a clear separation of the blood components. The erythrocytes aggregate and the granulocytes become slightly hypertonic, increasing their sedimentation rate, resulting in pelleting at the bottom of the ACCUSPIN tube. Lymphocytes and other mononuclear cells, i.e., monocytes, remain at the plasma-HISTOPAQUE-1077 interface.
Features and Benefits
ACCUSPIN™ System-Histopaque®-1077 offers numerous advantages, such as:
Optimal recovery of viable cells
Permits selective separation of blood cell types
Prevents cell distortion
Preserves cell viability
Minimal extraneous cell interference
Reproducible performance from batch to batch
Histopaque is sterile-filtered and endotoxin-tested
Stable at 2-8°C for at least 2 years when protected from light
Legal Information
Accuspin is a trademark of Sigma-Aldrich Co. LLC
Histopaque is a registered trademark of Merck KGaA, Darmstadt, Germany
