Hello everyone,
I am trying to get rid of hematopoietic lineage in bone marrow using the lineage depletion kit from Miltenyi. Without depletion, CD45-/Ter119- cells comprimise 8-10% of the whole population. After the depletion with the kit, this population was enriched only to 15-17%. In theory, with this kind of magnetic depletion kits efficiency should be really high so I was expecting to get rid of at least 70-80 percent of the hematopoietic cells. I used one LS column per 100 million cells so I dont think I saturated the column.
If anyone has an idea, I would be really glad to hear it :D
Thanks a lot in advance!
Nese
Hi Nese! One of my colleagues used the Lineage Cell Depletion Kit, which I am assuming is what you are using. He had no problem with his experiments. Keep in mind that magnetically-labelled antibodies are not stable for long periods (few months in fact). Also, you said you were using the LS columns. I don’t know how many cells you are using per column, but the protocol is written for 10 million cells. If you have more than 10 million cells, scale up the volumes. Other than that, I don’t know what the problem might be. Best of luck!
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