This is more of a cell biology question, but it's purpose is for use in flow assays, so...... During the course of in-vitro stimulation of both PBMCs and whole blood, we invariably see down regulation of certain surface antigens (for example, CD4 after 4 hours with PMA+Calcium Ionophore). These problems vary depending on stimulus and incubation times. This obviously creates problems as we try to identify populations following stimulation. This leads to two questions: 1. Is anyone aware if these antigens are actually down-regulated (i.e. internalized) or if they are shed into the supernatant? 2. As we use Brefeldin A to stop forward golgi transport (to trap cytokines within the cell for detection with fluorochrome conjugated Ab), would nocodazole, which blocks the return golgi pathway, work to inhibit the internalization of surface antigen? Thanks for any input you may have. Keith Bahjat Kbahjat@nwu.edu
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:55 EST