Re: acetone-extraceted liver powder /anti-luciferase
<< Previous Message | Next Message >>
From: | burch007@mc.duke.edu |
To: | Elizaha@aol.com |
Reply-To: | |
Content-Type: | text/plain; charset=us-ascii |
Elizabeth:
Years ago we use to absorb PoAb's with beef liver powder. It went like this:
Weigh out 100 mg liver powder per 1 ml antibody.
Place powder in a centrifuge tube.
Wash/hydrate the liver powder with PBS, vortex to mix.
Spin down in a powerful centrifuge. This will allow you to decant the PBS and
retain a hard pellet of liver powder.
Next add your antibody and break up the pellet. Use a wooden applicator stick or
whatever you can come up with. You can dilute your antibody 1:2 - 1:10 if the
volume is scarce and the working dilution titers out enough. Vortex the
liver/antibody mixture every 15 minutes or so over the course of an hour,
keeping it refrigerated or on ice. Centrifuge the mixture, recover your
antibody, re-titer to check the ab.
Hope this helps.
Regards,
Jim Burchette
Duke Immunopath
Durham NC
<< Previous Message | Next Message >>