5 replies to this topic

[bourkenm]

Hi,

Has anyone tried doing flow on PBMCs that have been in long term storage in liquid nitrogen? I am looking to both surface stain the PBMCs and also look at intracellular proteins (if that is possible).

Thanks,

N

[Angelie]

Yes, I have used very long term LN2 frozen PBMCs a no. of times for flow staining and it has worked pretty well both for surface staining and IC staining.

Angelie

bourkenm, on 11 October 2010 - 05:50 AM, said:

Hi,

Has anyone tried doing flow on PBMCs that have been in long term storage in liquid nitrogen? I am looking to both surface stain the PBMCs and also look at intracellular proteins (if that is possible).

Thanks,

N

[grayhair]

[quote name='Angelie' timestamp='1287590543' post='90062']
Yes, I have used very long term LN2 frozen PBMCs a no. of times for flow staining and it has worked pretty well both for surface staining and IC staining.

Angelie


[quote name='bourkenm' timestamp='1286805032' post='89227']
Hi,

Has anyone tried doing flow on PBMCs that have been in long term storage in liquid nitrogen? I am looking to both surface stain the PBMCs and also look at intracellular proteins (if that is possible).

Thanks,



Hi,

I have had cells in storage since 1992 and have done membrane as well as intracellular staining with great success. If the cells are keep
at around minus 135 you also can do functional assays on them!
Good Luck!

[bourkenm]

Great, thanks for the feedback!

[Denis Baev]

I am using LN frozen PBMCs also. It's good enough for phenotyping, subsetting, etc...But if your task is to catch Tregs by CD25high or to analyze CCR7 and other chemokine-Rs you have to use fresh spicemens ONLY.

Edited by Denis Baev, 19 January 2011 - 07:42 AM.

[HIV]

Angelie, on 20 October 2010 - 08:02 AM, said:

Yes, I have used very long term LN2 frozen PBMCs a no. of times for flow staining and it has worked pretty well both for surface staining and IC staining.


hi
can any one let me know what should be done for staining PBMC from human whole blood. I am trying to stain for various markers cd4, 8,19 extc.Its shows very poor staining, does any one has a clear cut protocol yu can share with.I am struggling with this problem since months

Thnx

Angelie

bourkenm, on 11 October 2010 - 05:50 AM, said:

Hi,

Has anyone tried doing flow on PBMCs that have been in long term storage in liquid nitrogen? I am looking to both surface stain the PBMCs and also look at intracellular proteins (if that is possible).

Thanks,

N