Hi everyone :)
I will be doing proteomic assay with blood from patients with leukemia. Is it necesary to isolate blast cells to reach a better proteomic result? or is it posible to just use ficol to separate lymphocytes (and keep normal and neoplastic in the mixture) and run the proteomic experiment and adjust the analysis later?
Thank you so much, I have been looking for this answer and I truly could not get It.
Hello Jorge.
As long as you anlayze control normal subject blood and patient blood together, it will be justified in the comparison. However, you are interested in the proteomes of cancerous cell in the blood of patient and normal cells of non-patient blood, you should isolate the cancerous cells.
Regards.
You should probably try to do cell sorting by flow cytometry after labelling leukemic cells with cell surface markers, e.g. CD19, CD10, CD22 for pre-B-ALL...!!!
Of course you need to separate the blast cells, especially using the peripheral blood, because blast cells are not always released from the bone marrow, or the proportion of the blast cells in blood are quite small. Therefore, blast cell purification is a must-do by either FACS sorting or CD19-microbead purification, and the same process shoule be done in parallel for controls .
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