IACF-Flow Cytometry Facility- Antibody Titration Protocol

Directly Coupled Antibody Titration Protocol

  1. In microtiter plate, serially dilute antibody in buffer.
  1. Serial Dilutions:

  1. Prepare cells for staining by resuspending them at 5x10^6/mL, and put 100uL of cells into test tubes (5x10^5 cells total).

  1. Add 10uL of appropriate antibody dilution to tubes according to the following schedule:

  1. Incubate for 20mins at 4C in the dark.

  1. Wash 2x in buffer.

  1. Analyze fresh or fix in 2% formaldehyde