How to increase IF signal in acetone/methanol fixed cells?
Posted 08 September 2009 - 09:58 AM
The results are the same from two different antibodies that work fine on Westernblots from the same cells and both antibodies are approved by the manufacturer (SCBT) for IF use. My target protein is a membrane-bound receptor so it should be present and is strongly positive by western with the same antibody.
Any ideas would be appreciated.
Posted 08 September 2009 - 04:46 PM
Posted 08 September 2009 - 06:29 PM
Try 2-4% paraformaldehyde (15 minutes @ room temp), wash, and then permabilize. 0.5% triton-x (5-15 minutes @ r.t.) does a great job with permabilization
Posted 11 September 2009 - 08:19 AM
Posted 14 September 2009 - 03:30 PM