Fixed Cell MLR

Palazzo, Adam AdamPa at BaylorHealth.edu
Mon Nov 28 15:27:22 EST 2005


Does anyone have any experience doing MLR with fixed APCs?  We're using 1% PFA for 20 mins to fix DCs after treatment with virus followed by 0.1M glycine to block reactive aldehyde groups.  Our T-cells proliferate for what looks like one generation but they are dead as seen with 7AAD staining when we analyze on the cytometer.  We're rinsing the DCs after fixation to ensure that no fixative is carrying over to the T-Cells.  If anyone has success with a particular protocol or method I would love to hear from you.   

Cheers,
Adam Palazzo
Baylor Institute for Immunology Research
Dallas, TX


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