Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum

Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum (http://www.molecularstation.com/forum/)
-   Biology Forum (http://www.molecularstation.com/forum/biology-forum/)
-   -   Western blot on FACS sorted Purkinje cells (http://www.molecularstation.com/forum/biology-forum/88196-western-blot-facs-sorted-purkinje-cells.html)

yvonne_GFP 03-10-2014 05:01 PM

Western blot on FACS sorted Purkinje cells
 
Hi,

We are trying to sort GFP Purkinje cells from all other cells in the cerebellum. We are having great trouble with this. Before going to the FACS facility we have 5 x 10^6 cells. The cells are passed through a 100 micrometer nozzle at a rate of 8000 events/ second. The sort decision has been based on both FSC, DAPI and GFP fluorescence. After the sort, which resulted in 800,000 cells, we have collected the cells in both media and PBS, spun the cells at 800rpm for 5 minutes and tried to run the samples on a Western blot and stain for GFP. We seem to be losing all GFP after the sort. Any help at all would be greatly appreciated.

Many thanks in advance,

Yvonne.


All times are GMT. The time now is 04:43 PM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved

Page generated in 0.08827 seconds with 11 queries