I am interested in gradient centrifugation of bone marrow cells that I could use to study gene and protein expression. I want to study neutrophil development in livestock. I understand this is worked out for humans but would like to know if there are any tips for non-humans.
Hi Jeffrey, I don't know if this may help you, but below is a link with lots of info about bone marrow cells isolation and gradient centrifugation. They discuss also on alternative protocols that may suit your needs as well.
https://www.researchgate.net/post/How_to_isolate_murine_Neutrophil_in_bone_marrow
Hi Jeffrey, I don't know if this may help you, but below is a link with lots of info about bone marrow cells isolation and gradient centrifugation. They discuss also on alternative protocols that may suit your needs as well.
https://www.researchgate.net/post/How_to_isolate_murine_Neutrophil_in_bone_marrow
If you know specific surface markers peculiar to the progenitor cells you are trying to isolate, why not develop fluorescent antibodies against those and FACS the cells? What quantities of cells would you like to isolate?
Yes, great idea. We have some antibodies which should work. I have paramagnetic beads which we have used to select cells. I need enough cells for gene expression. The presence of preformed protein in granules is likely the problem. We would like to be able to say "at stage 1, protein is already in granule and gene expresion is" We need to document when granule proteins come up, and stop being produced (stage). Thanks for you help.
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