Dear all, First, many thanks for all who have invested time to reply to my questions! I would like to summarize my conclusions from all flowers' answers: 1. Titration with small numbers of cells (1x10^6 cells) should be the first step, to find the optimal staining for the specific cells and antibody. The antibody should in excess and not the minimal amount that is adequate for good staining. 2. The Abs dilution is the critical point and shouldn't be changed in this kind of adjustment. 3. The cell number might be increased while keeping the same staining volume. The extent of increasing the cell number was not clear though, in some answers I got up to 5-fold increase and in some even more (about 50-fold). I would put it in the middle and say that increasing the cell number by 10-fold might still be reasonable. If anyone have any suggestions/additions, please write them. Hadi Aslan PhD, DMD Skeletal Biotech Lab/Prof. Dan Gazit The Hebrew University of Jerusalem- Hadassah Medical Center PO Box 12272, Jerusalem 91120 Israel Tel: 972-2-6757625 Fax: 972-2-6757628 Mobile: 972-545-909208 Web site: http://gazitlab.huji.ac.ilReceived on Thu Jun 26 19:58:00 2008
This archive was generated by hypermail 2.1.8 : Wed Jan 31 2007 - 03:12:00 EST
