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HELP: osteoclasts purification

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HELP: osteoclasts purification

Postby daylily » Sun Jun 17, 2007 10:09 pm

I want to get a pure populaiton of pre-osteoclasts from mouse bone marrow cells. I've read papers uing Sephadex G-10 column to purify bone marrow cells to get pure pre-osteoclasts. Does anybody know how this column work? How can it deplet other cell tpyes?
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Postby blcr11 » Mon Jun 18, 2007 8:22 am

I'd have to see the method. Are you sure you meant Sephadex G10? Sepahadex is a sizing resin. Its two primary uses are for clean up of protein preps usually as a final or near-final step, or separation of nucleotides and salts from a DNA or RNA reaction (e.g, when making labeled DNA) where such small molecules might be inhibitory for whatever next-steps are planned. The different grades of Sephadex have either larger or smaller pores that retain molecules up to a certain size. Anything larger passes around the resin and elutes from the column in the flow-through. I would think that all cells would be excluded from Sephadex (well, possibly not from G200, but even there I wouldn't think Sephadex could do much for resolving whole cells). There are modified Sephadex resins for ion exchange, but those would have names like DEAE-Sephadex, not Sephadex G10. So, I guess I'm skeptical that the method uses the G10 resin to separate cells, but I could be wrong.
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