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Cell Separation Using Magnetic Beads

Cell Separation Using Magnetic Beads. MiniMACS ® Separator. MS Column. Magnet. MACS ® Technology. MACS Technology Used For Cell Separation Magnetic Activated Cells Sorting Yields Relatively High Purity Cell Populations 3 Components Involved MACS MicroBeads MACS Column MACS Separator.

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Cell Separation Using Magnetic Beads

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  1. Cell Separation Using Magnetic Beads

  2. MiniMACS® Separator MS Column Magnet

  3. MACS® Technology • MACS Technology Used For Cell Separation • Magnetic Activated Cells Sorting • Yields Relatively High Purity Cell Populations • 3 Components Involved • MACS MicroBeads • MACS Column • MACS Separator

  4. MACS® Technology • MACS MicroBeads • 50 nm in size • Biodegradable • Conjugated to monoclonal antibodies • MACS Column • Generates strong magnetic field

  5. Labeling Of Cells With Beads Bead Monoclonal Ab

  6. Trapped Labeled Cells High Gradient Magnetic Field Magnetic Separation

  7. Cells Of Interest Magnetic Separation

  8. Direct Labeling • Advantages • Fast • Yields High Purity Cell Populations • Disadvantages • Binding of antibody to surface marker may activate cell • Some cell populations may require multiple markers to be identified

  9. Indirect Labeling

  10. Indirect Labeling • Useful in cases with dim surface marker expression • Allows for amplification of signal • Secondary antibody recognizes • Biotin • FITC • Fc portion of primary antibody • Streptavidin instead of secondary antibody • Recognizes biotin on primary antibody

  11. Depletion Strategy • Unwanted Cells Are Magnetically Labelled And Depleted • Useful In Following Cases • Removal of Unwanted Cells • No specific antibody is available for target cell • Binding of antibody to target cell results in activations

  12. Depletion Strategy

  13. Depletion Strategy

  14. Depletion Strategy

  15. Protocol For Isolation Of CD4+ Cells

  16. Protocol For Isolation Of CD4+ Cells

  17. Protocol For Isolation Of CD4+ Cells • 1. Prepare degased buffer

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