IGEM:Harvard/2007/Protocols/Indirect Magnetic Labeling Protocol


 * 1) After preparation of single-cell suspension, count cells, and centrifuge cell sample.
 * 2) Resuspend cell pellet and stain with the primary antibody according to the manufacturer's recommendations.
 * 3) For MACS Fluorochrome-conjugated or Biotinylated Antibodies, typically resuspend up to 10^7 total cells in 100 ul of buffer and add 10 ul of the respective antibodies.
 * 4) Mix well and refrigerate for 5-10 minutes or according to the manufacturer's recommendations. ##If fluorochrome-conjugated antibodies are used, incubate in the dark.
 * 5) Wash cells to remove unbound primary antibody by adding 1-2 mL of buffer per 10^7 cells and centrifuge at 300xg for 10 minutes.
 * 6) Repeat washing step
 * 7) Aspirate supernatant completely. Resuspend cell pellet in 80 ul of buffer per 10^7 total cells and add 20 ul of indirect MicroBeads per 10^7 total cells.
 * 8) Mix well and refrigerate for 15 minutes (4C)
 * 9) (Optional) When using unconjugated or biotinylated primary antibodies, cells can be fluorescently stained with a fluorochrome-conjugated antibody at this step.
 * 10) Wash cells by adding 1-2 mL of buffer per 10^7 cells and centrifuge at 300xg for 10 minutes. Aspirate supernatant completely.
 * 11) Resuspend up to 10^8 cells in 500 ul buffer.


 * 1) *Proceed to Magnetic Separation