Making Serial Antibody DilutionsReagents/Equipment: PBS or other appropriate buffer. Small capped tubes Pipettes capable of accurate delivery of 200 µL and 1000 µLvolumesKeep buffer and tubes in ice

1. Pipet 450 µL buffer into a tube.

2. Add 50 µL antibody solution and mix. This gives a 1:10 dilution of the antibody.

3. Label tubes A through M for 1:50, 1:100, 1:200, 1:400, etc. to 1:51,200 dilutions.

4. Pipet 1600 µL of dilution buffer into tube A (to become a 1:50 dilution). Pipette

1000 µL (1.0 mL) of dilution buffer into tubes B through M (to become 1:100–

1:102,400 dilutions).

5. Pipette 400 µL of 1:10 antibody dilution into tube A (which contains 1600 µL

buffer). Mix well. This results in a 1:50 antibody dilution.

6. Take 1000 µL of antibody sample from Tube A and add to Tube B (which contains

1000 µL buffer). Mix well.

7. Take 1000 µL of antibody sample from Tube B and add toTube C (which contains

1000 µL buffer), etc. Mix well.

A.C.R. Feb 2005