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Isolating Cells from Any Sample Source Without Columns

The following procedure provides guidelines for isolating human cells using EasySep™, a fast, easy and column-free cell isolation system capable of isolating virtually any cell type from any sample source.

This is a general procedure; specific conditions may vary according to the cell type being enriched. Find EasySep™ kits for a specific cell type by clicking here.

 

  1. Prepare the sample as a cell suspension in the recommended medium, at the concentration indicated on the product information sheet. (Cell Concentration Calculator)
  2. Add the EasySep™ cocktail at the concentration indicated on the product information sheet, mix well and incubate at room temperature (15 – 25°C).
  3. Vortex the magnetic particles. Ensure that particles are in a uniform suspension with no visible aggregates.
  4. Add the magnetic particles to your sample as indicated on the product information sheet. Mix well and incubate at room temperature (15 – 25°C).
  5. Bring the cell suspension up to the total volume recommended on the product information sheet by adding medium. Mix the cells in the tube by gently pipetting up and down 2 – 3 times. Place the tube (without cap) into the magnet. Set aside.
  6. The magnetically labeled cells will be held inside the tube, while the unlabeled cells can be poured off into a new tube. Separate the fractions as follows (depending on the EasySep™ magnet in use):
    • Pick up the EasySep™ magnet, and in one continuous motion invert the magnet and tube, pouring off the desired fraction into a new tube. Hold the magnet and tube in the inverted position for 2 – 3 seconds, and then return to upright position. Do not shake or blot off any drops that may remain hanging from the mouth of the tube.
    • Carefully pipette the unlabelled cell fraction into a new tube. Ensure that the pipettor does not touch the sides or the bottom of the original tube.

In negative selection, the desired cells are removed to the new tube. In positive selection, desired cells remain in the original tube. To calculate cell recovery after isolation, use the HIPapp Cell Recovery Calculator.

Have a question about this protocol/tip? Ask us.

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