Hi Joana,

I worked with this some years back. Although I worked with PBMCs, and not with spleen.

The procedure although lengthy, is nicely described in the information leaflet provided with the product. If you could tell me the problem that you are facing, I could try to help out.

There is another thread where you could get more info. (https://www.researchgate.net/post/Monocytes_isolation_from_blood)

Regards..

Spleen cells: You would need a single cells suspension from spleen before you go for enrichment using the kit. We generally use an artery clamp to squeeze the whole spleen in a dish with PBS and you can harvest maximum number of cells. Alternatively you can minse spleen and then mix rigorously with a fire-polished pasteru pipette using PBS. the latter would turn out to be messy.

Once you have the suspension, pass it through a filter, 30micron, to get single cell suspension, which would never be an absolute single-cell suspension though.

Using kit is really an easy go, its very clearly instructed in the leaflet.

To add to the above suggestion, another easy method to make single cell suspension from spleen is to keep it in between two glass slides and rub them together. This breaks the outer capsule and you get a suspension very easily.