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Human Platelets (For WB analysis)


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#1 scs

scs

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Posted 10 November 2010 - 04:00 PM

I had a request from a lab to isolate human platelets. Instead of generating a lysate form the platelets, the request was to just add electrophoresis buffer in a 1:1 ratio to run on WB to test primary antibodies. Here is my question:

1) Without lysing the platelets, would the platelet proteins just clump up on the top of a SDS-page gel and not correctly run since they are still contained in the cell fragment?
2) Without lysing the platelets and removing the cell debris, would not the platelets just collect at the bottom of a vial?

3) I believe the lab is wanting to test proteins within the thrombocytes and without lysing, the antibodies would not actively bind. Is this correct or is there something about the thrombocytes that I am overlooking.

Thanks for any help provided.

Edited by scs, 10 November 2010 - 04:03 PM.





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