I incubated 250´g of total lysate (at 1mg/mL) with 2.5-5 ´L antibody/IP (final concentration 10-20ul antibody/mg lysate) O/N at 4¯C. Then I added protein A agarose beads 2h at 4¯C. After 4 washes in RIPA buffer + protease inhibitors, elutions were done by addition of SLB2X and boil/vortex/boil/vortex.
Finally, when I did the western blot against Cpn10 with Ab108600 (1/5000 in 5%milk O/N at 4¯C) to observe the IP efficiency, I just got a really weak band of Cpn10 in my elutions in comparison with the 10% input. So, my IP yield was much less efficient than what I expected and not enough to be use in experiments.
We are sorry thatt his antibody did not work for this customer.
The antibody is covered by our Abpromise guarantee in both IP and Human, therefore the customer received a credit note in compensation.
We would like to encourage our customers to send us their feedback about the use of this antibody in IP. Please do not hesitate to contact our Scientific Support team for further information.
Abcam user community
Submitted Sep 29 2014
Get resources and offers direct to your inboxSign up