Shreffler:CFSE Labeling: Difference between revisions
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==CSFE Labeling== | ==CSFE Labeling== | ||
===Materials=== | |||
CFDA/SE [http://products.invitrogen.com:80/ivgn/en/US/adirect/invitrogen?cmd=catProductDetail&entryPoint=adirect&productID=C1157&messageType=catProductDetail Invitrogen C1157] as 5 mM stock in DMSO | CFDA/SE [http://products.invitrogen.com:80/ivgn/en/US/adirect/invitrogen?cmd=catProductDetail&entryPoint=adirect&productID=C1157&messageType=catProductDetail Invitrogen C1157] as 5 mM stock in DMSO | ||
===Procedure=== | |||
Suspend PBMCs at 10x10<sup>6</sup> cells/mL in PBS alone. | Suspend PBMCs at 10x10<sup>6</sup> cells/mL in PBS alone. | ||
Revision as of 18:38, 19 May 2009
CSFE Labeling
Materials
CFDA/SE Invitrogen C1157 as 5 mM stock in DMSO
Procedure
Suspend PBMCs at 10x106 cells/mL in PBS alone.
Cells should be in PBS for CFSE labeling.
If CFSE negative control is needed, remove cells now.
Make '2X' concentration (10 μM) in PBS
- Add 4mL PBS + 8 μL 5 mM CFSE (for example)
Combine 1:1 PBMC cells + CFSE (i.e. 300uL PBMCs + 300uL CFSE) in 15 ml tube.
Place in H2O bath x 10 min.
Wash in 10 mL complete medium (10 min at 300G). Aspirate.
Resuspend in medium at desired density
