Hello Berrin Saracoglu, 

What do you mean by recovery ? You can simply wash for several times with PBS  . What sort of experiment you are planning with your recovered blood ? 

Regards, 

Babu

Hi Babu;

I was adding bone marrow onto histopaque to isolate Mesenchymal stem cells. As you know you have to do it vey slowly not to mix the marrow and the ficoll. But Had an unlucky moment N I missed them .. dunno how many times I centrifuged it by PBS and tried to load on histopaque again but doest work anymore. no Buffy coat..

any suggestion?

simply dilute the entire sample three times in PBS, then centrifuged and recover the pellet, then finally back on ficoll.

thank you :)

Hi Berrin,

If still you have the problem, lyse the RBCs by using ACK lysing solution, after that wash twice with PBS. ACK lysin is used to Lyse the RBCs in the buffy layer.

Thanks.

thanks :)