[email protected] (DK) wrote in news:[email protected]:
> In article <[email protected]>, Dr > Engelbert Buxbaum <[email protected]> wrote: >>In article <[email protected]>, >>[email protected] says... >>> >>> Hi team - >>> >>> Anyone have a good protocol for isolating platelets and removing >>> plasma proteins through a Sepharose column? I've been having >>> trouble with this and fresh blood is hard to come by!!! >> >>You can not separate cell typess by gel filtration. This is usually >>done by centrifugation, either fractionated or in an isotonic >>gradient. Such gradients can be formed for example with ficoll, >>metrizamide or nycodenz. Since only cells, but not the plasma >>proteins, enter such a gradient, this step will also separate the >>cells from the proteins. > > I do believe platelets separation from plasma on Sepharose CL2B > is an established protocol. It's not clear what "trouble" above > actually means. Column clogging? Platelets lysing/activating? Low > yield? > > DK I forget the actual kind of Sepharose, but it is an established protocol to separate platelets from plasma. One can prepare platelet-rich plasma rather easily be choosing a) the right anticoagulant, b) discarding the first few drops or ml of blood so as to eliminate any "tissue juice" that can activate the platelets, and c) the right centrifugation conditions. I have never done Sepharose separation of plasma and platelets, but it is a well-known protocol. The difficulty is probably in knowing the exact volumes to use, and the right diameter tube. I'd contact Dr. Ellinor Peerschke at Mount Sinai in NY for experimental details. -- Best regards Han Broekman email address is invalid _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
