Serial Antibody Dilutions

Reagents/Equipment

PBS or other appropriate buffer

Small capped tubes

Pipets capable of accurate delivery of 200 mL and 1000 mL volumes

*Keep buffer and tubes on ice

Procedure

1. Pipet 450 uL buffer into a tube.
2. Add 50 uL 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 uL of dilution buffer into tube A (to become a 1:50 dilution). Pipette 1000 uL (1.0 mL) of dilution buffer into tubes B through M (to become 1:100 - 1:102,400 dilutions).
5. Pipette 400 uL of 1:10 antibody dilution into tube A (which contains 1600 uL buffer). Mix well. This results in a 1:50 antibody dilution.
6. Take 1000 uL of antibody sample from tube A and add to tube B (which contains 1000 uL buffer). Mix well.
7. Take 1000 uL of antibody sample from tube B and add to tube C (which contains 1000 uL buffer), etc. Mix well.

Serial Antibody Dilution Voumes