描述：
The BD IMag™ Human CD4 T Lymphocyte Enrichment Set - DM is used for the negative selection of CD4 T lymphocytes from peripheral
blood. The Biotinylated Human CD4 T Lymphocyte Enrichment Cocktail contains monoclonal antibodies that recognize antigens expressed
on erythrocytes, platelets, and peripheral leukocytes that are not CD4 T lymphocytes. The BD IMag™ Streptavidin Particles Plus - DM are
magnetic nanoparticles that have streptavidin covalently conjugated to their surfaces. With these two components, the BD IMag™ Human
CD4 T Lymphocyte Enrichment Set - DM avoids the inadvertent activation of the enriched CD4 T lymphocytes by using reagents that do not
directly bind to those T cells. This Enrichment Set has been optimized for use with the BD IMagnet™, and it contains sufficient reagents to
label 10^9 peripheral blood mononuclear cells (PBMC).
The detailed Magnetic Labeling and Enrichment Protocol follows. In summary, the Biotinylated Human CD4 T Lymphocyte Enrichment
Cocktail simultaneously stains erythrocytes, platelets, and most leukocytes except the CD4 T lymphocytes. After washing away excess
antibody, BD IMag™ Streptavidin Particles Plus - DM are added to the cell suspension and bind the cells bearing the biotinylated antibodies.
The tube containing this labeled cell suspension is then placed within the magnetic field of the BD IMagnet™ (Cat. No. 552311). Negative
selection is then performed to enrich for the unlabeled T cells. Labeled cells migrate toward the magnet (positive fraction), leaving the
unlabeled cells in suspension so they can be drawn off and retained (enriched fraction). The negative selection is repeated twice to increase the
yield of the enriched fraction. If greater purity is required, negative selection may be performed on the enriched fraction. For clarification of
the procedure, the magnetic separation steps are diagrammed in the Enrichment Flow Chart. The enriched (and positive) fractions can be
evaluated in downstream applications such as flow cytometry and tissue culture. The antibodies in the Biotinylated Human CD4 T
Lymphocyte Enrichment Cocktail have been optimized and pre-diluted to provide maximum efficiency for enrichment of CD4 T lymphocytes
from PBMC.
The CD4 T Lymphocyte Enrichment Cocktail is comprised from the following biotin-conjugated monoclonal antibodies:
Anti-human CD8, SK1
Anti-human CD11b/Mac-1 (CR3), clone ICRF44
Anti-human CD16, clone 3G8
Anti-human CD19, clone HIB19
Anti-human CD36, clone CB38 (NL07)
Anti-human CD56, clone B159
Anti-human CD123 (IL-3 Receptor α chain), clone 9F5
Anti-human CD235a (Glycophorin A), clone GA-R2 (HIR2)
Anti-human γδ TCR, clone B1
原厂资料：
The BD IMag™ Human CD4 T Lymphocyte Enrichment Set - DM is used for the negative selection of CD4 T lymphocytes from peripheral blood. The Biotinylated Human CD4 T Lymphocyte Enrichment Cocktail contains monoclonal antibodies that recognize antigens expressed on erythrocytes, platelets, and peripheral leukocytes that are not CD4 T lymphocytes. The BD IMag™ Streptavidin Particles Plus - DM are magnetic nanoparticles that have streptavidin covalently conjugated to their surfaces. With these two components, the BD IMag™ Human CD4 T Lymphocyte Enrichment Set - DM avoids the inadvertent activation of the enriched CD4 T lymphocytes by using reagents that do not directly bind to those T cells. This Enrichment Set has been optimized for use with the BD IMagnet™, and it contains sufficient reagents to label 10^9 peripheral blood mononuclear cells (PBMC).
The detailed Magnetic Labeling and Enrichment Protocol follows. In summary, the Biotinylated Human CD4 T Lymphocyte Enrichment Cocktail simultaneously stains erythrocytes, platelets, and most leukocytes except the CD4 T lymphocytes. After washing away excess antibody, BD IMag™ Streptavidin Particles Plus - DM are added to the cell suspension and bind the cells bearing the biotinylated antibodies. The tube containing this labeled cell suspension is then placed within the magnetic field of the BD IMagnet™ (Cat. No. 552311). Negative selection is then performed to enrich for the unlabeled T cells. Labeled cells migrate toward the magnet (positive fraction), leaving the unlabeled cells in suspension so they can be drawn off and retained (enriched fraction). The negative selection is repeated twice to increase the yield of the enriched fraction. If greater purity is required, negative selection may be performed on the enriched fraction. For clarification of the procedure, the magnetic separation steps are diagrammed in the Enrichment Flow Chart. The enriched (and positive) fractions can be evaluated in downstream applications such as flow cytometry and tissue culture. The antibodies in the Biotinylated Human CD4 T Lymphocyte Enrichment Cocktail have been optimized and pre-diluted to provide maximum efficiency for enrichment of CD4 T lymphocytes from PBMC.
The CD4 T Lymphocyte Enrichment Cocktail is comprised from the following biotin-conjugated monoclonal antibodies:
Anti-human CD8, SK1
Anti-human CD11b/Mac-1 (CR3), clone ICRF44
Anti-human CD16, clone 3G8
Anti-human CD19, clone HIB19
Anti-human CD36, clone CB38 (NL07)
Anti-human CD56, clone B159
Anti-human CD123 (IL-3 Receptor α chain), clone 9F5
Anti-human CD235a (Glycophorin A), clone GA-R2 (HIR2)
Anti-human γδ TCR, clone B1
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