Sample Purification

2.1.1. Sample Processing

The starting material is heparinised peripheral blood, which is centrifuged through a Histopaque (Sigma Diagnostics, UK) gradient to separate the lymphocytes and granulocytes.

1. 20 mL heparinised blood.

2. 50 mL conical based tubes (Bibby, UK).

3. Histopaque-1077 (Sigma Diagnostics, UK).

4. Plastic Pasteur pipets.

5. Phosphate buffered saline (PBS).

6. Deionised water.

7. Centrifuge to spin 50 mL tubes at 400g.

2.1.2. Fluorescent Assisted Cell Sorting (FACS)

1. Flow cytometer with sort facility plus associated equipment and consumables.

2. Mouse anti-human monoclonal antibodies specific for the tumor clone of interest (see Note 1).

2.1.3. Magnetic Microbead Purification

1. MACS CD19 microbeads (Miltenyi Biotec, Germany) plus associated equipment and consumables.

Was this article helpful?

0 0

Post a comment