[Cytometry] whole blood

Harvey, Shane HarveyS at Princeton.Huntingdon.com
Thu Feb 13 09:10:34 EST 2014


Barbara,

 

I have found that in the case of the lyse no wash protocol not working
that going back the lysing with ACK Lysis buffer does the trick. 

 

Stain blood with your antibodies of choice for 15-30min

Add 10:1 ACK/Blood volume of Lysis buffer

Lyse for 8-12 min, then spin down for 5min at 400g

Decant, Add your favorite buffer/1xPBS

Spin 5min at 400g, decant.

Add your Buffer/1xPBS Again, Spin for 5 min at 400g

Decant, Fix, and you are good to go. 

 

Shane Harvey 

Scientist

Huntingdon Life Sciences Inc
100 Mettlers Road 
Somerset NJ 08873, USA 

HarveyS at Princeton.Huntingdon.com
<mailto:HarveyS at Princeton.Huntingdon.com>  

 

 

 

 

 

Dear all,

I would like to know whether someone can suggest how to perform
immunofluorescence staining on whole blood (CD3, CD56, CD16, CD19,
CD14). I've tried to lyse red cells with BD FACS lysis solution but it
seems it doesn't work very well. Can anyone suggest a protocol to
efficiently lyse red cells?

Thanks a lot,

Barbara

 

Dr. Barbara Morandi

Immunology Unit

Istituto Nazionale per la Ricerca sul Cancro

CBA- L.go R.Benzi, 10

16132 - Genova, Italy

Tel :39 (010) 5558-221

Fax: 39 (010) 354282

E-mail: morandibarb at yahoo.it

 

 

 

 

 

 

 


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