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 MACS Column 50 nm in size
Biodegradable
Conjugated to monoclonal antibodies
MACS Column
Generates strong magnetic field

5. Labeling Of Cells With Beads
Monoclonal Ab

6. Magnetic Separation
High Gradient
Magnetic Field
Trapped Labeled Cells

7. Magnetic Separation
Cells Of Interest

8. Direct Labeling Advantages Disadvantages 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