Bio-Rad ReadyPrep™ 2-D Cleanup Kit - Manual (Page 10)

discard the supernatant. Centrifuge the tube briefly
(15–30 sec) and remove and discard any remaining
wash. The pellet will appear white at this stage. Air-dry
the pellet at room temperature for no more than 5 min
(the pellet will look translucent once sufficiently dry).
Note: Do not over-dry pellets. Over-dried pellets will be
difficult to resuspend.
13. Resuspend each pellet by adding an appropriate
volume of 2-D rehydration/sample buffer to the pellet.
Notes: Refer to Section 5.2 for recipes of typical 2-D
rehydration/sample buffers or Section 6 for a list of
convenient premade 2-D rehydration/sample buffers
available from Bio-Rad.
Consult Section 5.1 for a guide to selecting the volume of
2-D rehydration/sample buffer to use to resuspend the
protein pellet.
For SDS-PAGE, add an appropriate volume of Laemmli
sample buffer to the pellet, vortex the tube until the pellet
is dissolved and then heat the sample for 5 min at
95-100oC.
Vortex the tube for at least 30 sec. Incubate the tube
at room temperature for 3–5 min. Vortex the tube
again for ~1 min or pipet the solution up and down to
fully resuspend.
Notes: With some samples (large pellets or over-dry pellets)
the pellet may be difficult to resuspend. In these cases,
sonication can be used to speed the process.
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