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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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Cell separation using magnetic beads l.jpg

Cell Separation Using Magnetic Beads


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MiniMACS® Separator

MS Column

Magnet


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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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MACS® Technology

  • MACS MicroBeads

    • 50 nm in size

    • Biodegradable

    • Conjugated to monoclonal antibodies

  • MACS Column

    • Generates strong magnetic field


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Labeling Of Cells With Beads

Bead

Monoclonal Ab


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Trapped Labeled Cells

High Gradient

Magnetic Field

Magnetic Separation


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Cells Of Interest

Magnetic Separation


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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


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Indirect Labeling


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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


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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


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Depletion Strategy


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Depletion Strategy


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Depletion Strategy


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Protocol For Isolation Of CD4+ Cells


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Protocol For Isolation Of CD4+ Cells


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Protocol For Isolation Of CD4+ Cells

  • 1. Prepare degased buffer


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