producing antibodies takes some time. Am I able to separate intact T helper and T killer cells from each other and other blood cells without applying any antibody?
you can separate granulocytes and rbc from the rest of the WBC using a ficoll gradient. The protocol is easy and readily available on the web. After you harvest the WBC from the ficoll-media interface you can then separate T cells from B cells with a nylon wool column. The collected cells will contain T cells NK cells and monocytes. You can then attempt to remove the monocytes by 1 hour adherence to a petri dish. I am not aware of any method without antibodies to separate subsets of T cells or to separate NK cells from T cells.
let me know if you need more info on the nylon wool separation method - it's an old method but is very effective at removing B cells. there may be more updated methods than the one I used since it is more than 10 years since I did this.
Hi Bahram,
When you say 'without applying any antibody' I assume you don't want to label your cells of interest with an antibody for sorting, right?
There are diverse commercially available kits out there for so-called 'no touch' or negative selection of all types of peripheral blood immune cells. 'no touch' means that all cell types which you are not interested in are labeled with antibodies and then removed from your cell mix via a column. The cells of interest, e.g. CD4+ T cells, remain 'untouched' and are usually >95% pure. Have a look at the miltenyi website: http://www.miltenyibiotec.com/en/products-and-services/macs-cell-separation/cell-separation-reagents/t-cells/cd4-t-cell-isolation-kit-human.aspx
I hope this is helpful.
Christian
Hi Christian
Yes, I am trying to find a simple approach if it would be possible.
Thank you very much for your comments, they are so helpful.
Pooreydy
I was going to suggest the use of steel wool, but I see it has already been suggested
You can separate out a large percentage of adherent cells (monocytes) just by letting them sit on the bottom of a plastic well for a an hour or two in complete media with calcium and magnesium. Then gently wash out the non-adherent cells in PBS without calcium and magnesium. This will "enrich" for T & B cells.
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