Making Serial Antibody Dilutions Reagents/Equipment: PBS or other appropriate buffer.

Small capped tubes Pipettes capable of accurate delivery of 200 L and 1000 L volumes Keep buffer and tubes in ice 1. Pipet 450 L buffer into a tube. 2. Add 50 L 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 L of dilution buffer into tube A (to become a 1:50 dilution). Pipette 1000 L (1.0 mL) of dilution buffer into tubes B through M (to become 1:100 1:102,400 dilutions). 5. Pipette 400 L of 1:10 antibody dilution into tube A (which contains 1600 L buffer). Mix well. This results in a 1:50 antibody dilution. 6. Take 1000 L of antibody sample from Tube A and add to Tube B (which contains 1000 L buffer). Mix well. 7. Take 1000 L of antibody sample from Tube B and add toTube C (which contains 1000 L buffer), etc. Mix well.

A.C.R. Feb 2005