I'm currently trying to precipitate glycogen from platelets; however, the protocol I have uses ice cold 0.32 M sucrose buffer in platelet isolation process. I just can't figure out why?
That's an interesting question Ms Habibulla...
I think it's at least partly due to the high density of a sucrose solution. The increased density helps separate different components of cells (or platelets!) when you spin it.
Sucrose is also often used to stabilise cell and organelle membranes, but I'm not sure if this is of much assistance when you are using it in a lysis buffer...
All the best,
Paul
Thank you Paul. I tried using the ice-cold sucrose buffer as the protocol did. However, because of the sensitivity of the platelets to the cold, half of my pellet was lysed.
I've solved the problem. I used sonication plus boiling to release the glycogen and then acid hydrolysis them.
Thank you,
kind regards,
Mihray
Ice-cold sucrose solution would cause the activation and eventual death of platelets, releasing platelet components including glycogen. Cold itself can cause death of platelets. Perhaps concentration gradient due to sucrose in addition to the cold helps break the platelets effectively releasing platelet components including glycogen.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies