The RosetteSep™ Human NK Cell Enrichment Cocktail是通过阴选法从全血中分离NK细胞。全血中多余的非目标细胞是通过其四聚物的抗体复合物识别CD3、CD4、CD16、 CD36、CD56、CD66b、CD123及血型糖蛋白A等去除的。纯化的NK阳性细胞主要分布在血浆与浮力密度介质之间。
原厂资料:
Description
The RosetteSep™ Human NK Cell Enrichment Cocktail is designed to isolate NK cells from whole blood by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD3, CD4, CD19, CD36, CD66b, CD123 and glycophorin A on red blood cells (RBCs). When centrifuged over a buoyant density medium such as RosetteSep™ DM-L (Catalog #15705) or Lymphoprep™ (Catalog #07801), the unwanted cells pellet along with the RBCs. The purified NK cells are present as a highly enriched population at the interface between the plasma and the buoyant density medium.
Product Type:
Cell isolation products
Recommended For:
The isolation of human NK cells from whole blood by negative selection
Components:
RosetteSep™ Human NK Cell Enrichment Cocktail
Accessory Products:
• Lymphoprep™ (Catalog #07801), or
• RosetteSep™ DM-L (Catalog #15705)
Area of Interest:
Immunology
Cell Isolation Product Type:
Reagents
Cell Source:
Buffy coat, Whole blood
Cell Type:
NK cells
Popular Product Line:
RosetteSep™
Selection Method:
Negative
Species:
Human
Legal Statement:
Ficoll-Paque™ PLUS is a trademark of GE Healthcare Ltd.
Intended Use Statement:
FOR RESEARCH USE ONLY. NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES.
ISO Statement:
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485 MEDICAL DEVICE STANDARDS.
注意事项:
FAQS
HOW ROSETTESEP™ WORKS
Q. WHAT IS ROSETTESEP™?
A. RosetteSep™ is a rapid cell separation procedure for the isolation of purified cells directly from whole blood, without columns or magnets.
Q. HOW DOES ROSETTESEP™ WORK?
A. The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a density centrifugation medium (e.g. Ficoll-Paque™ PLUS, Lymphoprep™).
Q. WHAT FACTORS AFFECT CELL RECOVERY?
A. The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, density centrifugation medium, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the density centrifugation medium as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of density centrifugation medium can be collected without worry.
CELL TYPES AND SAMPLE SOURCES
Q. WHICH CELL SAMPLES CAN ROSETTESEP™ BE USED WITH?
A. RosetteSep™ can be used with leukapheresis samples, bone marrow or buffy coat, as long as: the concentration of cells does not exceed 5 x 107 per mL (can dilute if necessary); and there are at least 20 RBCs for every nucleated cell (RBCs can be added if necessary).
Q. CAN ROSETTESEP™ BE USED WITH PREVIOUSLY FROZEN OR CULTURED CELLS?
A. Yes. Cells should be re-suspended at 2 - 5 x 107 cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.
Q. CAN ROSETTESEP™ BE USED TO ENRICH PROGENITORS FROM CORD BLOOD?
A. Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll™, which can lead to a higher RBC contamination than peripheral blood separations.
Q. DOES ROSETTESEP™ WORK WITH MOUSE CELLS?
A. No, but we have developed Easy SepTM, a magnetic-based cell isolation system which works with mouse and other non-human species.
ANTI-COAGULANTS
Q. WHICH ANTICOAGULANT SHOULD BE USED WITH ROSETTESEP™?
A. Peripheral blood should be collected in heparinized Vacutainers. Cord blood should be collected in ACD.
Q. SHOULD THE ANTICOAGULANT BE WASHED OFF BEFORE USING ROSETTESEP™?
A. No, the antibody cocktail can be added directly to the sample.