Lymphocyte Isolation Medium
- Medium to separate lymphocytes from whole blood
For the isolation of lymphocytes from whole blood, this Lymphocyte Separation Medium can be used. It is a mixture of Ficoll® and sodium diatrizoate (Hypaque) with density adjusted to 1.077 g/ml. This sterile filtered product is intended for laboratory/manufacturing use, and is not for in vitro diagnostic use.
Lymphocyte Isolation Protocol
Isolation of lymphocytes from whole blood is very easy with Lymphocyte Separation Medium 1.077 and fits in general cell isolation protocols:
- Dilute whole blood 1:1 with calcium-magnesium-free PBS and layer 9 ml of diluted blood onto 6 ml Lymphocyte Separation Medium
- Centrifuge at 400 x g for 15 minutes
- Collect the interface by removing the top layer of plasma/PBS carefully. Dilute the interface in 20 ml serum-free medium
- Centrifuge at 70 x g for 10 minutes, discard supernatant and resuspend the pellet in 2-3 ml serum-free medium
- Count nucleated cells that include lymphocytes.
Lymphocytes will be concentrated at the interface (step 3), along with some platelets and monocytes.