Lidstrom:Ultracentrifuge

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(New page: Back to Protocols == Basics == * If you desire 4<sup>o</sup>C during your spin cycle, you must allow the machine to cool to that point before it will allow you to ...)
(Example Protocols)
Line 11: Line 11:
== Example Protocols ==
== Example Protocols ==
-
* To spin down the membranes and membrane-bound proteins in a crude extract, centrifuge for 100,000 RPM at 4<sup>o</sup>C for 1 hour.
+
* To spin down the membranes and membrane-bound proteins in a crude extract, centrifuge for 45,000 RPM at 4<sup>o</sup>C for 1 hour.  With our MLS-50 mushroom-shaped rotor, this is 163,000xg at r<sub>average</sub> (108,000xg at r<sub>minimum</sub>).

Revision as of 19:56, 26 May 2013

Back to Protocols

Basics

  • If you desire 4oC during your spin cycle, you must allow the machine to cool to that point before it will allow you to press start.
  • If you use too small of a volume in the ultracentrifuge tubes, they will collapse!
    • Tubes with only 1/2 mL liquid collapse in ultracentrifuge.
      Tubes with only 1/2 mL liquid collapse in ultracentrifuge.
    • It is hard to say how much is enough, but it seems to be about 1 mL. 1/2 mL definitely collapsed on Janet in 5/2013.
    • You probably won't be able to get any sample out of a collapsed tube.
    • When they collapse it is very tricky to get them out. Needle nose pliers are probably your best bet but it can still be very hard.
  • If the ultracentrifuge won't start and won't even try to start, open the door, lift the rotor up, place it back down, let it cool back down to your desired temp, and hold start.

Example Protocols

  • To spin down the membranes and membrane-bound proteins in a crude extract, centrifuge for 45,000 RPM at 4oC for 1 hour. With our MLS-50 mushroom-shaped rotor, this is 163,000xg at raverage (108,000xg at rminimum).
Personal tools