Coupling magnetic beads-antibody
Posted 13 February 2010 - 04:24 PM
I am washing three times my beads with PBS/BSA 5mg/ml before adding 2 to 7 microgram of antibody 2 hours or overnight at 4C on the wheel. Then, I wash twice my beads with PBS/BSA and add my lysate.
My IP didn't work and I detected my protein of interest in the flowthrough after the 2 hours or ON IP. I decided to test my beads and collect the flowthrough after the coupling beads-AB and found out that my antibody is there. I do not understand. I have done this in the past and worked really well. Do I miss something ?
Posted 17 February 2010 - 11:50 AM
Posted 18 February 2010 - 09:44 AM
Do you have any idea what is going on ?
Posted 18 February 2010 - 10:10 AM