How much whole blood should be used with 3 ml of Lymphocyte Separation Medium 1077 in a 15 ml conical tube?

For PBMC isolation using 3 ml of Lymphocyte Separation Medium 1077 (C-44010) in a 15 ml conical tube, we recommend loading 3-8 ml of diluted whole blood onto the separation medium.
Before layering, the whole blood should be diluted with PBS at a ratio of 1:2 or 1:3.
 
Example (1:2 dilution):

• Mix 1 volume of whole blood with 1 volume of PBS.
• To obtain 3-8 ml of diluted blood, use 1.5-4 ml of whole blood and add an equal volume of PBS.

As a general guideline:

• 15 ml conical tube: 3 ml Lymphocyte Separation Medium 1077 + 3-8 ml diluted whole blood.
• 50 ml conical tube: 15 ml Lymphocyte Separation Medium 1077 + 15-30 ml diluted whole blood.

For optimal separation, carefully layer the diluted blood onto the Lymphocyte Separation Medium without disturbing the interface.